LIBRARY 

Main  Lib, 
Agric,  Depfc 

UNIVERSITY  OF  CALIFORNIA. 


Deceive 

BIOLOGY 


Sl^/4  OGY 

Accessions  No.  1  '  {  (&<J    CLns  No. 


BACTERIOLOGY 

AND 

INFECTIVE      DISEASES 


A    TEXT-BOOK 


OF 


BACTEKIOLOGY 


INCLUDING   THE 


ETIOLOGY    AND    PREVENTION 


OF 


INFECTIVE    DISEASES 


AND  A   SHORT   ACCOUNT  OF 


YEASTS    AND    MOULDS,    ELEMATOZOA,    AND 
PSOROSPERMS 


OF  THE 

UNIVERSITY 
^^C^^H^/ 

BY 

EDGAE    M.    CEOOKSHANK,    M.B. 

PROFESSOR    OF   COMPARATIVE   PATHOLOGY   AND   BACTERIOLOGY,   AND   FELLOW   OF   KING** 
COLLEGE,    LONDON. 


FOURTH  EDITION 
RECONSTRUCTED,    REVISED    AND    GREATLY    ENLARGED 


PHILADELPHIA 

W.    B.    SAUNDEES 

925  WALNUT  STREET 
1897 


BlOLO^f 

LIBRARY 

G 


UN 


7/7^3 


SIR    JOSEPH    LISTER,    BART.,    M.B.,    P.R.S., 

WHO    HAS   CREATED   A   NEW   EPOCH    IN 

MEDICINE    AND    SURGERY, 

BY    APPLYING    A    KNOWLEDGE    OF    MICRO-ORGANISMS 
TO    THE     TREATMENT    OF    DISEASE, 

l&h'vs  Silorh  is,  totth  permission,  JHpefoiratefc 
BY    THE    AUTHOR 

AS    A 
TOKEN    OF    ADMIRATION    AND    RESPECT 


PREFACE 


TO  THE 


FOURTH    EDITION. 


THIS  book,  though  nominally  a  fourth  edition,  is  practically 
speaking  a  new  work.  The  progress  of  Bacteriology  has  been 
very  rapid,  and  many  new  investigations  have  been  made  in 
connection  with  the  etiology,  prevention  and  treatment  of 
communicable  diseases.  It  has  been  necessary  to  reconstruct, 
enlarge  and  thoroughly  revise  the  text  of  the  third  edition, 
and  I  have  added  twenty-six  chapters. 

The  most  important  researches  conducted  in  bacteriological 
laboratories  are  those  relating  to  the  contagia.  In  many 
diseases  of  man  and  animals  it  has  not  been  possible  to 
identify  the  contagium  with  a  bacterium,  or  indeed  with  any 
micro-organism ;  but  when  the  virus  is  chemically  examined, 
or  investigated  with  a  view  to  protective  inoculation,  or  utilised 
for  experiments  in  serum-therapeutics,  such  researches  are 
within  the  province  of  the  bacteriologist. 

The  recognition  of  the  fact  that  in  so  many  diseases  the 
nature  of  the  contagium  has  not  yet  been  determined  will 
have  the  effect  of  encouraging  continued  activity  in  this 
important  field  of  scientific  investigation. 

I  hope  that  this  work  will  continue  to  be  of  use  as  a  text- 
book for  the  bacteriological  laboratory,  and  that  the  chapters 
on  the  etiology  and  prevention  of  the  communicable  diseases 


viii  PREFACE   TO   THE   FOURTH   EDITION. 

of  man  and  animals  will  be  not  only  of  scientific  interest, 
but  of  practical  value  to  Medical  Officers  of  Health  and 
Veterinary  Inspectors. 

I  have  divided  the  book  into  three  parts.  Part  I.  is 
mainly  technical,  and  includes  the  most  recent  methods 
employed  in  studying  bacteria  and  investigating  the  etiology 
of  disease.  Part  II.  deals  with  infective  diseases  and  the 
bacteria  associated  with  them.  Any  clinical  or  pathological 
evidence  which  may  help  to  throw  light  on  the  nature  and 
origin  of  the  contagia  is  taken  into  account.  The  most 
effectual  measures  for  stamping  out  these  diseases  are 
referred  to,  as  they  are  intimately  connected  with  a 
knowledge  of  the  life-history  of  micro-organisms.  Part  III. 
contains]  descriptions  of  about  five  hundred  bacteria.  Many 
are  of  no  practical  importance  and  of  very  little  scientific- 
interest,  but  a  text-book  for  the  laboratory  cannot  be  con- 
sidered complete  unless  an  account  is  given  of  all  bacteria 
which  have  been  more  or  less  completely  investigated.  I 
have  endeavoured  to  refer  to  the  original  descriptions  and 
to  verify  them  by  comparison  with  actual  cultivations,  but 
in  a  very  great  number  of  instances  this  has  been  quite 
impossible,  and  I  desire  to  acknowledge  the  assistance  I  have 
received  from  the  works  of  several  authors,  especially  those 
of  Fliigge,  Fraukel,  Eisenberg,  Baumgarten,  Frankland, 
Sternberg,  Lehmann  and  Neumann. 

If^have  rearranged  the  bibliography  according  to  the 
chapters^and  the  names  of  authors  are  given  in  alphabetical 
order.  With  the  aid  of  the  current  numbers  of  the  Annales 
<le  rinstitut  Pasteur,  the  Zeitschrift  fur  Hygiene,  the 
Centralblatt  fur  Bakteriologie  und  Parasitenkunde^  and  the 
Journal  of  Comparative  Pathology  and  Bacteriology,  it  is 
possible  to  become  acquainted  with  the  most  recent  litera- 
ture of  the  subject. 

Many  of  the  coloured  plates  illustrating  the  last  edition 
have  not  been  reproduced.  Those  substituted  for  them  have 
been  drawn  from  my  own  preparations,  and  most  of  them 


PREFACE   TO   THE    FOURTH    EDITION.  IX 

have  already  appeared  in  my  Reports  to  the  Board  of 
Agriculture  and  in  other  publications. 

One  hundred  and  thirty-three  woodcuts  and  photographs 
have  been  added  in  the  text,  and  I  have  reverted  to  the  plan 
which.  J  adopted  in  the  second  edition,  of  having  many  of 
them  printed  in  colours. 

I  take  this  opportunity  of  thanking  Professor  Frankel  for 
kindly  permitting  me  to  reproduce  some  of  the  photographs 
in  his  excellent  Atlas. 

I  ana  particularly  indebted  to  Professor  Hamilton  for 
the  use  of  cliches  of  figures  in  his  classical  treatise  on 
Pathology j  and  to  the  New  Sydenham  Society  for  several 
from  the  English  translation  of  Professor  Fliigge's  well- 
known  work  on  micro-organisms. 

To  my  Demonstrator,  Dr.  George  Newman,  D.P.H.,  I  am 
indebted  for  much  assistance  in  correcting  the  proof-sheets, 
and  for  the  preparation  of  an  index. 

EDGAR  M.  CROOKSHANK. 

SAINT  HILL,  EAST  GEINSTEAD.  SUSSEX, 
August  1st,  1896. 

P.S. — Since  this  work  was  finally  passed  for  press  the  con- 
clusions of  the  Royal  Vaccination  Commissioners-  have  been 
published.  I  have  at  the  last  moment  added  extracts  in  the 
form  of  a  supplementary  Appendix.  E.  M.  C. 

September  IStJt,  1896. 


CONTENTS. 


PAET     I. 

THEORETICAL    AND    TECHNICAL. 

PAGE 

CHAPTER    I. 

HISTORICAL   INTRODUCTION .         1 

CHAPTER    II. 

MORPHOLOGY   AND    PHYSIOLOGY    OF   BACTERIA  .  .  .       1 1 

CHAPTER    III. 

EFFECT    OF    ANTISEPTICS   AND    DISINFECTANTS    ON    BACTERIA       .       30 

CHAPTER    IV. 
CHEMICAL   PRODUCTS   OF   BACTERIA 39 

CHAPTER    V. 

IMMUNITY  .  .  .  .  .  .  .       '     .  .49 

CHAPTER    VI. 

ANTITOXINS   AND   SERUM   THERAPY 56 

CHAPTER    VII. 
THE   BACTERIOLOGICAL   MICROSCOPE 65 

CHAPTER    VIII. 
MICROSCOPICAL   EXAMINATION   OF   BACTERIA    .  .  .  .83 

CHAPTER    IX. 

PREPARATION  OF  NUTRIENT  MEDIA   AND   METHODS   OF   CULTIVA- 
TION .  99 


Xii  CONTENTS. 

PAGE 

CHAPTER    X. 

EXPERIMENTS   UPON   THE    LIVING   ANIMAL         .  .  .134 

CHAPTER    XI. 
EXAMINATION   OF   AIR,    SOIL,    AND   WATER         .  .  .  .    140  ,/ 

CHAPTER    XII. 

PHOTOGRAPHY    OF   BACTERIA  '.  .  .  .  .  .150 


PAET     II. 

ETIOLOGY   AND    PREVENTION    OF   INFECTIVE 
DISEASES. 

CHAPTER    XIII. 
SUPPURATION,    PY^MIA,    SEPTIOEMIA,    ERYSIPELAS    .  .  .173 

CHAPTER    XIV. 
ANTHRAX 191 

CHAPTER    XV. 

QUARTER-EVIL. — MALIGNANT  CEDEMA. — RAG-PICKERS'  SEPTI- 
CAEMIA.— SEPTICAEMIA  OF  GUINEA-PIGS. — SEPTICAEMIA  OF 
MICE  .  .  217 

CHAPTER    XVI. 

SEPTICAEMIA  OF  BUFFALOES.—  SEPTIC  PLEURO-PNEUMONIA  OF 
CALVES. — SWINE  FEVER. — SEPTICAEMIA  OF  DEER. — SEPTI- 
CAEMIA OF  RABBITS. — FOWL  CHOLERA. — FOWL  ENTERITIS. 
—DUCK  CHOLERA.— GROUSE  DISEASE  .  .  .  .226 

CHAPTER    XVII. 

PNEUMONIA. — INFECTIOUS    PLEURO-PNEUMONIA    OF    CATTLE.—- 

INFLUENZA 233 

CHAPTER    XVIII. 
ORIENTAL     PLAGUE. — RELAPSING     FEVER. — TYPHUS     FEVER.— 

YELLOW   FEVER    .  .250 


CONTENTS.  Xlll 

CHAPTER    XIX. 

SCARLET   FEVER. — MEASLES 261 

CHAPTER    XX. 

SMALL-POX.— CATTLE   PLAGUE        .  .  ...    284 

CHAPTER    XXI. 

SHEEP-POX. — FOOT-AND-MOUTH    DISEASE  ....    297 

CHAPTER    XXII. 

HORSE-POX. — COW-POX  .  .  .  .  .  .  .303 

CHAPTER    XXIII. 
DIPHTHERIA  .........    330 

CHAPTER    XXIV. 

TYPHOID   FEVER  .........    340 

CHAPTER    XXV. 

SWINE   FEVER       .  .  .  .  .  .  .  .  .347 

CHAPTER    XXVI. 

SWINE  MEASLES. — DISTEMPER  IN  DOGS.— EPIDEMIC   DISEASE   OF 

FERRETS.— EPIDEMIC    DISEASE    OF   MICE  .  .  .  .355 

CHAPTER    XXVII. 

ASIATIC  CHOLERA. — CHOLERA  NOSTRAS. — CHOLERAIC  DIARRHCEA 
FROM  MEAT  POISONING.  —  DYSENTERY. — CHOLERAIC  DIAR- 
RHCEA IN  FOWLS 360 

CHAPTER    XXVIII. 

TUBERCULOSIS .  .  .375 

CHAPTER    XXIX. 

LEPROSY. — SYPHILIS. — RHINOSCLEROM A.— TRACHOMA        .  .    406 

CHAPTER    XXX. 

ACTINOMYCOSIS. — MADURA   DISEASE          .  .413 

CHAPTER    XXXI. 

GLANDERS  .  .451 


xiv  CONTENTS. 

PAGE 

CHAPTER    XXXII. 
TETANUS. — RABIES.— LOUPING-ILL 457 

CHAPTER    XXXIII. 
FOOT-ROT •  464 

CHAPTER    XXXIV. 

FOUL-BROOD. — INFECTIOUS  DISEASE  OF  BEES  IN  ITALY. — 
PE"BRINE. — FLACHERIE. — INFECTIOUS  DISEASE  OF  CATER- 
PILLARS .469 


PAET      III. 

SYSTEMATIC   AND    DESCRIPTIVE. 

CHAPTER    XXXV. 

CLASSIFICATION   AND   DESCRIPTION   OF   SPECIES  .    475 


APPENDICES. 

APPENDIX    I. 

YEASTS  AND   MOULDS  .  .  .  .  .  .  .  .    577 

APPENDIX    II. 

ILEMATOZOA   IN   MAN,    BIRDS,   AND    TURTLES. — ILEMATOZOA    IN 

EQUINES,    CAMELS   AND   FISH. — ILEMATOZOA   IN   FROGS        .    589 

APPENDIX    III. 

PSOROSPERMS   OR   COCCIDIA. — AMCEBA   COLI      ....    609 

APPENDIX    IV. 

APPARATUS,   MATERIAL    AND    REAGENTS    EMPLOYED   IN   A   BAC- 
TERIOLOGICAL  LABORATORY  .  .  .  .  .  .612 

APPENDIX    V. 
BIBLIOGRAPHY 639 

SUPPLEMENTARY    APPENDIX. 

EXTRACTS   FROM   THE   FINAL   REPORT   OF   THE   ROYAL    VACCINA- 
TION   COMMISSION  .    667 


LIST     OF     ILLUSTRATIONS. 


WOOD   ENGRAVINGS   AND   PHOTOGRAPHS. 

FIG.  PAGE 

1.  Ascococcus  Billrothii,  x  65  (Cobn)         .  .  .  .  .14 

2.  Spirochgeta  from  Sewage  Water,  x  1200  (E.M.C.)         .  .       15 
'6.  Flagella  (Koch,  Brefeld,  Warming,  Zopf)            .                                      .16 

4.  Bacillus  Megatherium  (De  Bary)     .         .  .  .  .  .17 

5.  Clostridium  Butyricum,   x  1020  (Prazmowski)  .  .  .  .18 

6.  Leuconostoc   Meseuteroides ;    Cocci-chains  with   Arthrospores  (Van 

Tieghem  and  Cienkowski)        .  .  .  .  .  .19 

7.  Spore-bearing  Threads  of    Bacillus   Anthracis,  double-stained   with 

Fucbsine  and  Metbylene  Blue,  x   1200  (E.M.C.)       .  .  .20 

8.  Bacilli  of  Tubercle  in  Sputum,  x  2500  (E.M.C.)  .       21 

9.  Comma  Bacilli  in  Sewage  Water,  stained  with  Gentian  Violet,  x  1200 

(E.M.C.)  ........       22 

10.  Vibrios  in  Water  contaminated  with  Sewage,  x   1200  (E.M.C.)  .       22 

11.  Refraction  of  Light  (Carpenter)      ....  .66 

12.  Spherical  Aberration  (Carpenter)  .  .  .  .  .67 

13.  Combination   of  Lenses  in  Abbe's   Homogeneous  Immersion  (Car- 

penter) .........       67 

14.  Chromatic  Aberration  (Carpenter)  .  .  .  .  .68 

15.  Objective  with  Collar  Correction  (Zeiss)  .  .69 

16.  Microscope— English  Model  (Swift)        .  71 

17.  Removable  Mechanical  Stage  (Swift)     .  .  .  .  .72 

18.  Microscope — Continental  Model  (Zeiss)  .  .  .  .  .78 

19.  Iris  Diaphragm  (Zeiss)     ...  74 
20. .  Abbe's  Condenser  (Zeiss)  .......       75 

21.  Microscope  Lamp  (Baker)  .  .    ,  .  .  .6 

22.  Large  Microscope  Lamp  (Swift)  .  ,  .  .  .  .77 

23.  Arrangement  of  Powell  and  Lealand's  Microscope  in  working  directly 

on  the  Edge  of  the  Flame,  with  Stand  for  Micrometer  Eye-piece  to 
secure  Steadiness  and  Accuracy  of  Measurement  (Carpenter  after 
Nelson)  .  .  .  .  .  ,  .  .  .79 

24.  Ramsden  Micrometer  Eye-piece  (Swift)  .  80 
_.").   Micrometer  Eye-piece  (Zeiss)       .             .            .             .             .             .81 

26.  Inoculating  Needles  (E.M.C.)       .  .       84 

27.  1   eezing  Microtome  (Swift)  .  .94 
L's.  Microtome  (Jung)              .                         .  .95 

29.  Wire-cage  for  Test-tubes  (Muencke)        .....     100 

30.  Hot-air  Steriliser  (E.M.C.)  .     101 


xvi  LIST   OF   ILLUSTRATIONS. 

FIG.  PAGE 

31.  Hot-water  Filtering  Apparatus  (Muencke)  .  102 

32.  Method  of  making  a  Folded  Filter  (E.M.C.)       .  .  103 

33.  Steam  Steriliser  (Baird  and  Tatlock)      .  .  103 

34.  Incubator  (Muencke)        .  •  104 

35.  Method  of  Inoculating  a  Test-tube  containing  Sterile  Nutrient  Jelly 

(E.M.C.)  ....  .105 

36.  Levelling  Apparatus  (E.M.C)       .  .     107 

37.  Iron  box  for  Glass  Plates  (Muencke)       .  .     108 

38.  Method  of  Inoculating  Test-tubes  in  the  Preparation  of  Plate-cultiva- 

tions (E.M.C.)  .  •     108 

39.  Damp-chamber  containing  Plate-cultivations  (E.M.C)  .             .110 

40.  Pasteur's  Large  Incubator  (Becker)        .  .111 

41.  Petri's  Dish  (Becker)         .             .  .112 

42.  Glass  Benches  and  Slides  (Becker)          .  .112 

43.  Koch's  Serum  Steriliser  (Muencke)          ...  .     114 

44.  Hueppe's  Serum  Inspissator  (Baird  and  Tatlock)  .     115 

45.  Box  for  Sterilising  Instruments  (Becker)             .  116 

46.  Damp  Chamber  for  Potato-cultivations  (E.M.C.)            .  .            .     117 

47.  Apparatus  for   Sterilisation  by  Steam   under  pressure  (Baird  and 

Tatlock)             ..'...  .119 

48.  Drop  Cultivation  (Fltigge)            .             .            .             .            .  .121 

49.  Simple  Method  of  forming  a  Moist  Cell  (Schafer)          .            .  .122 

50.  Warm  Stage  (Schafer)      .....  .123 

51.  Warm  Stage  shown  in  Operation  (Schafer)         .  .     123 

52.  Warming  Apparatus  in  Operation  (Isiael)  ....     124 

53.  Section  of  Warming  Apparatus  and  Drop-culture  Slide  (Israel)  .     125 

54.  Israel's  Warming  Apparatus         ....  •     125 

55.  Gas  Chamber  in   use  with  Apparatus  lor  generating  Carbonic  Acid 

(Schafer) 126 

56.  Gas  Chamber  (Schafer)     .  .  .  .  .  .  .126 

57.  Moist  Cell  adapted  for  Transmission  of  Electricity  (Schafer)   .  .     127 

58.  Apparatus  arranged  for  Transmitting  Electricity  (Schafer)      .  .     127 

59.  Slide  with  Gold-leaf  Electrodes  (Schafer)  .  ,     128 

60.  Lister's  Flask  (Becker)     .  .  .  .  128 

61.  Sternbergs  Bulb  (Becker)  .  .  128 

62.  Aitken's  Tube  (Becker)     .  .  .129 

63.  Miguel's  Bulb  (Becker)     ....  129 

64.  Pasteur's  Flask  (Baird  and  Tatlock)        .  .  .130 

65.  Pasteur's  Double  Tube  (Baird  and  Tatlock)         .  .130 

66.  Frankel's  Anaerobic  Tube-culture  (Frankland)  .  .     131 

67.  Anaerobic  Culture  Tube  (Liborius)          .  .  .  .132 

68.  Apparatus  for  Anaerobic  Cultures  (Roscoe  and  Lunt)  .  .     133 

69.  Koch's  Syringe  (Baird  and  Tatlock)         .  .  135 

70.  Syringe  with  Asbestos  Plug  (Baird  and  Tatlock)  .  .135 

71.  Hesse's  Apparatus  (Muencke)       ......     142 

72.  Sedgwick  and  Tucker's  Tube  (Baird  and  Tatlock)  .  .  .     143 

73.  Pouchet's  Aeroscope  (Hamilton)  .....     143 

74.  Apparatus  for  Estimating  the  number  '_£  Colonies  in  a  Plate-cultiva- 

tion (Muencke)  .  .  .  .  .  .  .  146 

75.  Esmarch's  Roll-culture  (Frankland)        .....  147 

76.  Apparatus  for  Counting  Colonies  in  a  Roll-culture  (Becker)     .  .  148 

77.  Horizontal  Micro-photographic  Apparatus  (Swift)         .  .  .  15(5 


LIST   OF   ILLUSTRATIONS.  XV11 

Ki<;  PAGE 

78.  Reversible  Micro-photographic  Apparatus  (E.M.C.)     .  .  .     157 

79.  Reversible  Micro-photographic  Apparatus  arranged  in  the  Vertical 

Position  (E.M.C.)  .  .  .158 

80.  Large  Micro-photographic  Apparatus  (Swift)  ....     160 

81.  Photograph  of  an  Impression  Preparation  (E.M.C.)      .  .  .     162 
*2.  Photograph  of  a  Cultivation  of  Bacillus  anthracis  (E.M.C.)    .             .     168 
*:J.  Suppuration  of  Subcutaneous  Tissue  (Cornil  and  Ranvier)      .  .     174 
M.  Pus  with  Staphylococci,  x  800  (Fltigge)          .                                     .177 

85.  Subcutaneous  Tissue  of  a  Rabbit  forty-eight  hours  after  an  Injection 

of  Staphylococci,  x  950  (Baumgarten)  ....     177 

86.  Ulcerative  P^ndocarditis  :  Section  of  Cardiac  Muscle,  x    700  (Koch)     183 

87.  Pure-cultures  of  Streptococcus  Pyogenes  (E.M.C.)      .  .  .     184 

88.  Section  of  Skin  in  Erysipelas  (Cornil  and  Ranvier)      .  .  .185 

89.  Streptococcus  Pyogenes  Hominis  ;  Pure-cultures  on  Nutrient  Gela- 

tine (E.M.C.)    .187 

90.  Streptococcus  Pyogenes  Bovis  ;  Pure-cultures  on  Nutrient  Gelatine, 

(E.M.C.)  ....  .     188 

91.  Gonococcus,   x  800  (Bumm)       ......     190 

92.  Bacillus  Anthracis,  x  1200.    Blood  Corpuscles  and  Bacilli  unstained ; 

from  an  Inoculated  Mouse  (Frankel  and  Pfeiffer)     .             .             .  192 

93.  Pure-cultivation  of  Bacillus  anthracis  in  Nutrient  Gelatine  (E.M.C.)  193 

94.  Colonies  of  Bacillus  anthracis,   x  86  (Flttgge)             .            .            .  194 

95.  Impression -preparation  of  a  Colony,   x   70  (E.M.C.)     .            .            .  194 

96.  Margin  of  a  Colony,  x  250  (E.M.C.) 195 

97.  Filaments  with  Oval  and  Irregular  Elements,  x   800  (E.M.C.)           .  195 

98.  Spores  of  Bacillus  anthracis  stained  with  Gentian  Violet,    x    1500 

(E.M.C.) 197 

99.  Anthrax  in  Swine  (E.M.C.) 203 

100.  Anthrax  in  Swine  (E.M.C.)         .  .  .     205 

101.  Bacilli  of  Quarter-evil,  x   1000  (Frankel  and  Pfeiffer)  .  .     218 

102.  Pure-culture   of    Bacilli   of   Quarter-evil    in   Grape-sugar    Gelatine 

(Frankel  and  Pfeiffer)  .  .  .  .  .  .  .218 

103.  Bacilli  of  Malignant  (Edema,  x  950  (Baumgarten)      .  .  .     221 

104.  Pure-culture   of    Bacillus   of    Malignant   (Edema    in    Grape-Sugar 

Gelatine  (Frankel  and  Pfeiffer)  .  .  .  .  .222 

105.  Bacilli  of  Malignant  (Edema,   x  1000  (Frankel  and  Pfeiffer)  .     223 

106.  Pure-cultivation  of  the  Bacillus  of  Septicaemia  of  Mice  in  Nutrient 

Gelatine  (E.M.C.)          .  .  .  .  .  .  .225 

107.  Bacterium  of  Rabbit  Septicaemia  ;  Blood  of  Sparrow,  x  700  (Koch)     228 

108.  Bacterium  of  Fowl-cholera,  x  1200  (E.M.C.)  .  ,  .  .228 

109.  Bacterium  of  Fowl-cholera,  x  2500  (E.M.C.)  .  .228 

110.  Bacterium  of  Fowl-cholera;    Section   from  Liver  of  Fowl,    x    700 

(Fltigge)  ...  .     229 

111.  Bacillus  of  Hsemorrhagic  Septicaemia,  x  950  (Baumgarten)  .  .     231 

112.  Bacillus   of    Haemorrhagic   Septicaemia:    Pure-culture   in   Gelatine 

(Baumgarten)    .  .  .  .  .  .  .  •     231 

113.  Bacterium  Pneumonire  Crouposre  from  Pleural  Cavity  of  a  Mouse, 

x   1500  (Zopf)  ...  .234 

114.  Frieclliinder's    Pneumococcus ;    Pure-culture   in   Nutrient    Gelatine 

(Baumgarten)    ........     234 

115.  Capsule  Cocci  from  Pneumonia,  x   1500  (Baumgarten)  .     235 

116.  Micrococcus  of  Sputum  Septicaemia,  x   10CO  (Frankel  and  Pfeiffer)  .     23f> 

b 


xviii  LIST   OF   ILLUSTRATIONS. 

FIG.  PACK 

117.  Colonies  of  Sternberg's  Micrococcus,  x  100  (Frankel  and  Pfeiffer)    .  237 

118.  Acute  Catarrhal  Pneumonia,  x  480  (Hamilton)            >            .            .  239 

119.  Infectious  Pleuro-pneumonia  of  Cattle  (Hamilton)      .            .            .  240 

120.  Infectious  Pleuro-pneumonia  of  Cattle  (Hamilton)      .  .  .241 

121.  Bacillus  of  Influenza,  x  1000  (Itzerott  and  Niemann)            .            .  248 

122.  Bacillus  of  Influenza,  x  1200  (E.M.C)                                                    .  249 

123.  Bacilli  of  Plague  and  Phagocytes,  x  800  (Aoyama)    .             .            .  253 

124.  Spirillum  Obermeieri  in  Blood  of  Monkey  inoculated  with  Spirilla 

after  Removal  of  the  Spleen  (Soudakewitch)             .            .             .  258 

125.  Pure-cultivations  of  Streptococcus  Pyogenes  (E.M.C.)                         .  263 

126.  Free  Surface  of  Diphtheritic  Larynx,  x  350  (Hamilton)        .            .  331 

127.  Bacillus  of  Diphtheria  ;  from  a  Cultivation  on  Blood  Serum,  x  1000 

(Frankel  and  Pfeiffer) .  .  .  .  .  .  .332 

128.  Pure-cultures  of  Bacillus  Diphtheria  on  Glycerine-Gelatine  (E.M.C.)  333 

129.  Typhoid  Fever.     Ileum  of  Adult,  showing  Sloughy  and  Infiltrated 

Patches  (Hamilton)      .  .  .  .  .  .  .341 

130.  Typhoid    Bacilli  from    a    Colony   on    Nutrient  Gelatine,    x    1000 

(Frankel  and  Pfeiffer)  .  .  .  .  .  .  .342 

131.  Typhoid  Bacilli,  x  950  (Baumgarten)  .....  342 

132.  Flagella  of  Typhoid  Bacilli,  x   1000  (Frankel  and  Pfeiffer)     .            .  343 

133.  Colonies  of  the  Typhoid  Bacillus  (Frankel  and  Pfeiffer)          .             .  344 

134.  Pure-culture  of  Typhoid  Bacilli  inoculated  in  the  Depth  of  Nutrient 

Gelatine  (Baumgarten)    .                     .            .            .            .            .  344 

135.  Typhoid  Bacilli  in  a  Section  of  Spleen,   x  800  (Fliigge)         .             .  345 

136.  Typhoid  Bacilli  in  a  Section  of  Intestine  invading  the  Submucous 

and  Muscular  Layers,  x   950  (Baumgarten)  ....  346 

137.  Ulceration  of  the  Intestine  in  a  Typical  Case  of  Swine-fever  (E.M.C.)  348 

138.  Klein's  Bacillus  of  Swine-fever  (No.  1)             .            .            .             .  349 

139.  From  a  Preparation   of   Bronchial   Mucus  of  Klein's  Swine-fever 

Bacillus  (No.  2)             .......  349 

140.  Bacilli  from  an  Artificial  Culture  with  Spores,  Bacillus  No.  2  (Klein)  349 

141.  Blood  of  Fresh  Spleen  of  a  Mouse  after  Inoculation  with  Swine-fever 

Bacillus  No.  2  (Klein) 350 

142.  Bacilli  of  Swine  Erysipelas  (Baumgarten)        ....  356 

143.  Blood  of  Pigeon  inoculated  with  Bacilli  of  Swine  Erysipelas,  x  600 

(Schutz)  .  .  ...  .  .  •  .356 

144.  Pure-culture  in  Nutrient  Gelatine  of  Bacilli  from  Swine  Erysipelas 

(Baumgarten)    .                         .            .            .            .            .            .  357 

145.  Cover-glass  Preparation  of  a  Drop  of  Meat  Infusion  containing  a 

Pure-cultivation  of  Comma-bacilli  (Koch)      .  .  .  .361 

146.  Arthrospores  of  Comma-bacilli  (Hueppe)          ....  361 

147.  Flagella  of  Comma-bacilli  ;  stained  by  Loffler's  Method  (Frankel 

and  Pfeiffer)     .            .            .            .             .            .            .            .  :$62 

148.  Involution  Forms  of  Comma-bacilli,  x  700  (Van  Ermengem)             .  362 

149.  Colonies  of  Comma-bacilli  on  Nutrient  Gelatine  ;  natural  size  (Koch)  362 

150.  Colonies  of  Koch's  Comma-bacilli,  x  60  (E.M.C.)        .            .            .  363 

151.  Cover-glass  Preparation  from  the  Contents  of  a  Cholera  Intestine, 

x  600  (Koch)    .  .  .  .  .  .  .363 

152.  Cover-glass  Preparation  of  Cholera  Dejecta  on  Damp  Linen,  x  600 

(Koch)  .........  363 

15.'!.  Section  of  the   Mucous  Membrane  of  a  Cholera  Intestine,    x    600 

(Koch)   .            .                                                                                       .  364 


LIST   OF   ILLUSTRATIONS.  XIX 

FIG.  PAGE 

154.  Pure-cultivations  in  Nutrient  Gelatine  of  Koch's  and  of  Finkler's 

Comma-bacilli  (E.M.C.)  ......     365 

155.  Comma-shaped   Organisms    with    other    Bacteria  in    Sewage-con- 

taminated Water,   x   1200         ......  36(5 

156.  Comma-bacilli  of  the  Mouth,  x  700  (Van  Ermengem)  .  .  367 

157.  Finkler's  Comma-bacilli,  from  Cholera  nostras,  x  700  (Fliigge)        .  367 

158.  Deneke's  Comma-bacilli,  from  Cheese,  x  700  (Fliigge)  .  .  367 

159.  Pure-cultivation  of  the  Spirillum  Finkler- Prior,  in  Nutrient  Gelatine 

(E.M.C.)  .  .  .  .  .  ...  .370 

160.  Tropical  Dysentery ;  Mucous  Membrane  of  Large  Intestine  (Hamilton)  372 

161.  Tubercle  of  the  Lung  in  a  very  Early  Stage,  x  400  (Hamilton)         .  376 

162.  Primary  Tubercle  of  Lung  two  to  three  weeks  old,  x  50  (Hamilton)  377 

163.  Large  Oval  Giant  Cell  from  Tubercle  of  Lung,  x  300  (Hamilton)     .  377 

164.  Bacillus  Tuberculosis,  from  Tubercular  Sputum,   x  2500  (E.M.C.)     .  379 

165.  Pure-cultivation  of  the   Tubercle  Bacillus  on  Glycerine  Agar-agar 

(E.M.C.) .     380 

166.  Pure-cultivation  in  Glycerine  Agar-agar  after  ten  months'  growth 

(E.M.C.)  .  ,  ...  .  .381 

167.  Pure-cultivations  of  Tubercle  Bacillus  in  Glycerine  Agar-agar ;  a  sub- 

culture from  a  Pure-culture  in  Glycerine  Milk  (E.M.C.)  .            .  381 

16S.  Section  through  a  Lupus  Nodule  of  the  Nose  (Hamilton)  .            .  387 

169.  Tubercular  Ulceration  of  Mucosa  of  Ileum  (Hamilton)  .            .  388 

170.  Section   of  Lupus  of  the    Skin ;    Giant  Cell   containing  Tubercle 

Bacillus  (Fliigge)          .......     389 

171.  Tuberculosis  of  Pleura;  "Grape-disease"  (E.M.C.)      .  .  .390 

172.  Tubercular  Ulceration  of  the  Intestine  of  a  Cow  (E.M.C.)      .  .     393 
1 ::!.  Tubercular  Ulceration  of  the  Intestine  of  a  Rabbit  (E.M.C.)  .  .     395 

174.  Tubercular  Lungs  of  Rabbit  (E.M.C.)    .  .  .  .  .396 

175.  Cover-glass  Preparation  of  Pus  from  a  Chancre,  x  1050  (Lustgarten)     410 
17<).  Wandering  Cell  containing  Bacilli  (Lustgarten)  .  .  .     410 

177.  Section  of  Liver  from  a  Case  of  Actinomycosis  in  Man  (E.M.C.)         .  417 

178.  Actinomycotic  Tumour  in  the  Throat  of  a  Steer  (E.M.C)         .             .  424 

179.  Actinomycotic  Tumour  of  the  Cheek  (E.M.C.)  ....  424 

180.  Steer  with  Emaciation  the  Result  of  Actinomycosis  (E.M.C.)             .  425 

181.  Actinomycotic    Growths    from    the    Pleura    resembling    "  Grape- 

Disease"  (E.M.C.)         .  .  .  .  .  .425 

182.  Actinomycosis  of  the  Skin  (E.M.C.)       .            .  .  .  .430 
is:!.  Part  of  Human  Foot  with  Madura  Disease  (E.M.C.)  .  .  .448 
1S4.  Bacilli  of  Glanders,   x   700  (Fliigge)      .            .  .  .  .452 

185.  Section  of  a  Branch  of  the  Pulmonary  Artery  showing  Glanders 

Bacilli  penetrating  the  Wall  (Hamilton)        ....     453 

186.  Pure-culture  of  the  Tetanus  Bacillus  in  Grape-sugar  Gelatine  (Frankel 

and  Pfeiffer)      .  .  .  .  .  .  .  .458 

187.  Foot  of  Sheep  showing  Disease  of  Horn  (Brown)         .  .  .     465 

188.  Section  through  the  Foot  showing  a  Crack  extending  through  the 

Wall 465 

189.  Secreting  Membrane  covered  with  Fungoid  Growths  (Brown)  .     466 

190.  Advanced  Form  of  Disease  of  Skin  between  the  Claws  (Brown)  .     466 

191.  Distortion  of  Hoof  in  an  Advanced  Form  of  Foot-rot  (Brown)  .     467 

192.  Diseased  Comb  (Cowan)              .            .            .            .            .  .469 

193.  Spores  of  Bacillus  Alvei  (E.M.C.)           .            .            .             .  .470 
191.  Pure-culture  in  Nutrient  Gelatine  (Cheshire  and  Cheyne)      .  .     470 


XX  LIST   OF  ILLUSTRATIONS. 

*-:u.  rAOK 

195.  Cultivation  on  the  Surface  of  Gelatine  (Cheshire  and  Cheyne)           .  471 

196.  Cladothrix  Dichotoma  (Zopf)     .            .                                                  .  479 

197.  Friedlander's  Pneumococcus,  x   1500  (Zopf)    .            .                         .  483 

198.  Ascococcus  Billrothh  (Cohn)      .                                                               .  498 

199.  Clostridium  Butyricum  (Prazmowski)   .            .            .            .            .  503 

200.  Bacillus  Cyanogenus,  x  650  (Neelsen)             ....  507 
"201.  Pure-cultivation  of  Bacillus  figurans  on  the   Surface  of  Nutrient 

Agar-agar  (E.M.C.)       .            .                                                               .  509 

202.  Photograph   of    Part   of  an  Impression    Preparation    of    Bacillus 

figurans  on  Nutrient  Gelatine,  x  50  (E.M.C.)            .            .            .  510 

203.  Part  of  the  same  Specimen,  x  200        .  .  .  .  .510 

204.  Bacillus  Indicus  :  Colonies  in  Agar,  x  60        .  .  .  .518 

205.  Bacillus  Neapolitans,  x  700  (Emmerich)        ....  522 

206.  Bacillus  Megatherium  (De  Bary)  .  .  .  .  .524 

207.  Pure-culture  of  Bacillus  Megatherium  in  Gelatine  (E.M.C.)   .             .  524 

208.  Bacillus  Putrificus  Coli,  x  1000  (Bienstock)    ....  529 

209.  Bacillus  Pyogenes  Foetidus,  x  790  (Passet)     ....  530 

210.  Bacillus  Saprogenes,  No.  1  (Rosenbach)            .                                      .  531 

211.  Bacillus  Subtilis  with  Spores  (Baumgarten)     ....  535 

212.  Pure-culture  of  Bacillus  Subtilis  in  Nutrient  Gelatine  (Baumgarten)  .  535 

213.  Pure-culture  of  Bacillus  Subtilis  on  the  Surface  of  Nutrient  Agar 

(E.M.C)             .                                                                                       .  536 

214.  Bacterium  Zopfii  (Kurth)           ......  542 

215.  Beggiatoa  Alba  (Zopf)     .....  .543 

216.  Phase-forms  of  Beggiatoa  Persicina  (Warming)           .            .             .  544 

217.  Cladothrix  Dichotoma  (Zopf)     ......  545 

218.  Crenothrix  Kuhniana  (Zopf)      ......  546 

219.  Leuconostoc  Mesenteroides  (Van  Tieghem  and  Cienkowski)  .            .  550 

220.  Micrococcus  in  Pyaemia  in  Babbits  (Koch)       ....  556 

221.  Proteus  Mirabilis  ;  Swarming  Islands  on  the  Surface  of  Gelatine, 

x  285  (Hauser) 5(51 

222.  Proteus  Mirabilis  ;  Involution  Forms,   x   524  (Hauser)           .            .  5(51 

223.  Proteus  Vulgaris,  x  285  (Hauser^          .  .  .  .  .562 

224.  Sarcina,  x  600  (Fliigge)             ......  5(53 

225.  Spirochteta  Plicatile  (E.M.C.)  .             .             .             .             .             .  566 

226.  Comma-bacilli  in  Water  contaminated  with  Sewage  .            .             .  567 

227.  Comma-bacilli  of  the  Mouth,  x  700  (Van  Ermengem)             .            .  568 

228.  Deneke's  Comma-bacilli,  from  Cheese,  x  700  (Fliigge)           .            .  568 

229.  Streptococcus  in  Progressive  Tissue  Necrosis  in  Mice  (Koch)             .  571 

230.  Vibrio  Rugula,  x  1020  (Prazmowski)    .  .  .  .  .574 

231.  Black  Torula;  Pure-cultivation  on  Potato  (E.M.C.)     .            .            .  581 

232.  Head  and  Neck  of  Calf  with  Advanced  Ringworm  (Brown)  .            .  585 

233.  Non-pigmented  Amoeboid  Forms  (Marchiafava  and  Celli)     .            .  590 

234.  Pigmented  Amoeboid  Forms  (Golgi)      .....  590 

235.  Semi-lunar  Bodies  of  Laveran  (Golgi)  ...                         .  590 

236.  Rosette  Forms  with  Segmentation  (Golgi)     .   .                                     .  591 

237.  Flagellated  Forms  (Vandyke  Carter)     .  .  .  .  .591 

238.  "  Surra  "  Parasites,  occurring  Singly  and  Fused,  x   1200         .            .  598 

239.  Parasites  in  the  Blood  of  Rats  (Lewis)              ....  599 

240.  A  Monad  in  Rat's  Blood,  x   3000  (E.M.C.)       ....  601 

241.  Monads  in  Rat's  Blood,  x  1200  (E.M.C.)         ....  602 

242.  Monads  in  Rat's  Blood  stained  with  Methyl  Violet,  x   1200  (E.M.C.)  603 


LIST   OF   ILLUSTRATIONS.  XXI 

HO.  I'AGK 

243.  Organisms  in  the  Blood  of  Mud-fish  (Mitrophanow)   .  .  .     604 

244.  Organisms  in  the  Blood  of  the  Carp  (Mitrophanow)    .  .  .     605 

245.  Amoeba  cola  in  Intestinal  Mucus  (Losch)          ....     611 

2  It;.  Warm  Stage  (Schafer)    .  .  .  .  .  .  .613 

247.  Warm  Stage  (Strieker)  ...  .     614 

IMS.  Combined  Gas  Chamber  and  Warm  Stage  (Strieker)    .  .  .     614 

249.  Vertical  Micro-photographic  Apparatus  (Leitz)  .  .  .     622 

250.  Koch's  Steam  Steriliser  (Muencke)        .....     623 

251.  Hot-air  Steriliser  (Muencke)      ......     623 

252.  Section  of  Hot-air  Steriliser  (Muencke)  .  .     623 

253.  Hot-water  Filtering  Apparatus  with  Ring  Burner  (Rohrbeck)  .     624 

254.  Wire-cage  for  Test-tubes  (Muencke)      ,  626 

255.  Platinum-needles  (E.M.C.)        ......     626 

256.  Damp  Chamber  for  Plate-cultivations  (E.M.C.)  .  .  .626 

257.  Apparatus  for  Plate  cultivations  (E.M.C.)         .  .  .627 
253.  Box  for  Glass-plates  (Muencke)              .             .  .627 

259.  Glass  Benches  for  Glass-plates  (Becker)           ....  627 

260.  Israel's  Case  (Becker)      .                         .                                                   .  628 

261.  Damp  Chamber  for  Potato  Cultivations  (E.M.C.)         .                         .  628 

262.  Koch's  Serum  Steriliser  (Muencke)        .                                                  .  629- 

263.  Serum  Inspissator  (Muencke)     ....                         .  62D 

264.  D'Arsonval's  Incubator  (Muencke)                                  ,  631 

265.  Schlosing's  Membrane  Regulator  (Muencke)    ....  632 

266.  Gas  Burner  protected  with  Mica  Cylinder  (Muencke)                           .  633 

267.  Koch's  Safety  Burner  (Muencke)           .....  633 

268.  Babes'  Incubator  (Muencke)      .  .  .  .634 

269.  Moitessier's  Gas-pressure  Regulator  (Muencke)             .             .             .  634 

270.  Reichert's  Thermo-regulator  (Muencke)            .                                      .  635 

271.  Meyer's  Thermo-regulator  (Muencke)    .....  636 

272.  Siphon  Bottle  with  Flexible  Tube,  Glass  Nozzle,  and  a  Mohr's  Pinch- 

cock  (E.M.C.)    .637 

273.  Desiccator  (E.M.C.)         .  .  .  .  .  .  .638 


DESCRIPTION     OP     PLATES. 


DESCRIPTION    OF    PLATE    I. 

Bacteria,  Schizomycetes,  or  Fission  Fungi. 

Following  p.  14. 

1.  Cocci  singly  and  varying  in  size.  2.  Cocci  in  chains  or  rosaries  (strepto- 
coccus). 3.  Cocci  in  a  mass  (staphylococcus).  4  and  5.  Cocci  in  pairs 
(diplococcus).  6.  Cocci  in  groups  of  four  (merismopedia).  7.  Cocci  in  packets 
(sarcina).  8.  Bacterium  termo.  9.  Bacterium  termo  x  4000  (Dallinger.and 
Drysdale).  10.  Bacterium  septiccemice  hcemorrhagicce .  11.  Bacterium  pneu- 
monice  crouposce.  12.  Bacillus  subtilis.  13.  Bacillus  murisepticus.  14. 
Bacillus  diphtheria.  15.  Bacillus  typhosus  (Eberth).  16.  Spirillum  undula 
(Cohn).  17.  Spirillum  volutans  (Cohn).  18.  Spirillum  cholera  Asiaticce. 
19.  Spirillum  Obermeieri  (Koch).  20.  Spirochceta  plicatilis  (Fliigge).  21. 
Vibrio  rugula  (Prazmowski).  22.  Cladothrix  Forsteri  (Cohn).  23.  Cladothrix 
dichotoma  (Cohn).  24.  Nonas  OJtenii  (Cohn).  25.  Nonas  Warmingii  (Cohn). 
26.  Rhabdomonas  rosea  (Cohn).  27.  Spore-formation  (Bacillus  alvei).  28. 
Spore-formation  (Bacillus  anthracis).  29.  Spore-formation  in  bacilli  cultivated 
from  a  rotten  melon  (Frankel  and  Pfeiffer).  .30.  Spore-formation  in  bacilli 
cultivated  from  earth  (Frankel  and  Pfeiffer).  31.  Involution-form  of  Crenothrix 
(Zopf).  32.  Involution-forms  of  Vibrio  serpem  (Warming).  33.  Involution- 
forms  of  Vibrio  rugula  (Warming).  34.  Involution-forms  of  Clostridium 
polymyxa  (after  Prazmowski).  35.  Involution-forms  of  Spirillum  cholera 
AsiaticfB.  36.  Involution-forms  of  Bacterium  aceti  (Zopf  and  Hansen). 
37.  Spirulina-form  of  Beggiatoa  alba  (Zopf).  38.  Various  thread-forms  of 
Bacterium  merismopedioides  (Zopf).  39.  False-branching  of  Cladothrix^Zopf). 


DESCRIPTION    OF    PLATE    II. 

Pure-cultivations  of  Bacteria. 

Following  p.  100. 

FIG.  1. — In  the  depth  of  Nutrient  Gelatine.  A  pure-cultivation  of  Koch's 
comma-bacillus  (Spirillum  choleras  Asiaticae)  showing  in  the  track  of 
the  needle  a  funnel-shaped  area  of  liquefaction  enclosing  an  air-bubble, 
and  a  white  thread.  Similar  appearances  are  produced  in  cultivations  of 
the  comma-bacillus  of  Metchnikoff. 

FIG.  2. —  On  tlie  surface  of  Nutrient  Gelatine.  A  pure-cultivation  of  Bacillus 
typhosus  on  the  surface  of  obliquely  solidified  nutrient  gelatine. 

xxii 


DESCRIPTION  OF  PLATES.  xxiii 

FIG.  3. —  On  the  surface  of  Nutrient  Agar-agar.  Pure-cultivation  of  Bacillus 
indicus  on  the  surface  of  obliquely  solidified  nutrient  agar-agar.  The 
growth  has  the  colour  of  red  sealing-wax,  and  a  peculiar  crinkled 
appearance.  After  some  days  it  loses  its  bright  colour  and  becomes 
purplish,  like  an  old  cultivation  of  Micrococcus  prodigiosus. 

FIG.  4. — On  the  surface  of  Nutrient  Agar-agar.  A  pure-cultivation  obtained 
from  an  abscess  (Staphylococcus  pyogenes  aureus). 

FIG.  5.—  On  the  surface  of  Nutrient  Agar-agar.  A  pure-cultivation  obtained 
from  green  pus  (Bacillus  pyocyaneus).  The  growth  forms  a  whitish, 
transparent  layer,  composed  of  slender  bacilli,  and  the  green  pigment 
is  diffused  throughout  the  nutrient  jelly.  The  growth  appears  green  by 
transmitted  light,  owing  to  the  colour  of  the  jelly  behind  it. 

FIG.  (\.-On  tlie  surface  of  Potato.  A  pure-cultivation  of  the  bacillus  of 
glanders  on  the  surface  of  sterilised  potato. 


DESCRIPTION    OF    PLATE    III. 

Plate-cultivation. 

Following  p.  108. 

This  represents  the  appearance  of  a  plate-cultivation  of  the  comma-bacillus 
of  Cholera  nostras,  when  it  is  examined  over  a  slab  of  blackened  plate-glass. 
The  drawing  was  made  from  a  typical  result  of  thinning  out  the  colonies  by 
the  process  of  plate-cultivation.  At  this  stage  they  were  completely  isolated 
one  from  the  other ;  but  later  they  became  confluent,  and  produced  complete 
liquefaction  of  the  gelatine. 

DESCRIPTION    OF    PLATE    IV. 

Streptococcus  Pyogenes. 

Following  p.  178. 

FIG.  1. — From  a  cover-glass  preparation  of  pus  from  a  pyaemic  abscess. 
Stained  with  gentian-violet  by  the  method  of  Gram,  and  contrast-stained 
with  eosin.  x  1200.  Powell  and  Lealand's  apochromatic  T\  Horn.  imm. 
E.  P.  10. 

FIG.  2. — From  cover-glass  preparations  of  artificial  cultivations  of  the  strepto- 
coccus in  broth  and  in  milk  at  different  stages  of  growth,  x  1200.  Powell 
and  Lealand's  apocbromatic  -fa  Horn.  imm.  E.  P.  10. 

In  these  preparations  there  is  a  great  diversity  in  size  and  form  of  the 
chains  and  their  component  elements.  In  the  drawing  examples  are 
figured  of  the  following: 

(a)  Branched  chains. 

(b)  Simple   chains    composed   of  elements   much   smaller  than  the 
average  size. 

O)  Chains  with  spherical  and  spindle-shaped  elements  at  irregular 

intervals.     These  are  conspicuous  by  their  size,  and  are  sometimes 

terminal. 

(d  e)  Chains  in  which  the  elements  are  more  or  less  uniform  in  size. 
(/)  Complex  chains  with  elements  dividing  both  longitudinally  and 

transversely,  and  varying  considerably  in  size  in  different  lengths 

of  the  same  chain. 


XXiv  DESCRIPTION   OF   PLATES. 

DESCRIPTION    OF    PLATE    V. 

Bacillus  Anthracis. 

Following  p.  192. 

FIG.  1. — From  a  cover-glass  preparation  of  blood  from  the  spleen  of  a  guinea- 
pig  inoculated  with  blood  from  a  sow.  x  1200.  Powell  and  Lealand's 
apochromatic  T\  Horn.  imm.  E.  P.  10. 

FIG.  2. — From  a  section  of  a  kidney  of  a  mouse.  Under  a  low  power  the 
preparation  has  exactly  the  appearance  of  an  injected  specimen.  Under 
higher  amplification  the  bacilli  are  seen  to  have  threaded  their  way  along 
the  capillaries  between  the  tubules,  and  to  have  collected  in  masses  in 
the  glomeruli.  Stained  with  Gram's  method  (gentian-violet),  and  eosin. 
x  500. 

FIG.  3. — Bacillus  anthracis  and  Microcoocus  tetragenits.  From  a  section  from 
the  lungs  of  a  mouse  which  had  been  inoculated  with  anthrax  three  days 
after  inoculation  with  Mlcrococcus  tetragenus.  A  double  or  mixed  infection 
resulted.  Anthrax-bacilli  occurred  in  vast  numbers,  completely  filling  the 
small  vessels  and  capillaries,  and  in  addition  there  were  great  numbers 
of  tetrads.  Stained  by  Gram's  method  (gentian- violet),  and  with  eosin. 
x  500. 


DESCRIPTION    OF    PLATE    VI. 

f 

Bacillus  Murisepticus. 

Following  p.  224. 

FIG.  1. — From  a  section  of  a  kidney  of  a  mouse  which  had  died  after  inocula 
tion  with  a  pure-cultivation  of  the  bacillus.     With  moderate  amplification, 
the   white  blood-corpuscles  have  a  granular  appearance,  and  irregular 
granular  masses  are  scattered  between  the  kidney  tubules.     Stained  by 
Gram's  method  with  eosin.      x  200. 

FIG.  2. — Part  of  the  same  preparation  with  high  amplification.  The  granular 
appearances  are  found  to  be  due  to  the  presence  of  great  numbers  of 
extremely  minute  bacilli,  x  1500. 


DESCRIPTION    OF    PLATE    VII. 

Casual  Cow-pox. 

Following  p.  278. 

FIG.  1. — Case  of  W.  P ,  a  milker,  infected  from  the  teats  of  a  cow  with 

natural  cow-pox.  There  was  a  large  depressed  vesicle  with  a  small 
central  crust  and  a  tumid  margin,  the  whole  being  surrounded  by  a 
well-marked  areola  and  considerable  surrounding  induration. 

FIG.  2. — The  same  case  a  week  later,  showing  a  reddish-brown  crust  on  a 
reddened  elevated  and  indurated  base. 


DESCRIPTION   OF    PLATES.  XXV 

DESCRIPTION  OF  PLATE  VIII. 

Bacillus  diphtherias  and  Bacillus  typhosus. 

Following  p.  382. 

Kid.  1. — Cover-glass  preparation  from  a  pure-cultivation  of  Bacillus  diph- 
therias on  blood  serum  ;  obtained  from  the  throat  in  a  typical  case  of 
diphtheria.  Stained  with  gentian-violet,  x  1200. 

FIG.  2. — Cover-glass  preparation  from  a  pure-cultivation  of  Bacillus  typhosus 
on  nutri,ent-agar ;  from  the  spleen  in  a  case  of  typhoid  fever,  Stained 
with  gentian-violet,  x  1200. 


DESCRIPTION    OF    PLATES    IX.    AND    X. 

Swine  Fever. 

Following  p.  348. 

PLATE  IX. — Part  of  intestine  from  a  typical  case  of  swine  fever,   showing 

scattered  ulcers  and  ulceration  of  the  ileo-csecal  valve. 
PLATE  X. — From  the  same  case  of  swine  fever.    The  lungs  were  extensively 

inflamed  and  partly  consolidated,  and  the  lymphatic  glands  were  enlarged 

and  of  a  deep  reel  or  reddish-purple  colour. 


DESCRIPTION    OF    PLATE    XI. 

Bacillus  tuberculosis. 

Following  p.  378. 

The  figures  in  this  plate  represent  the  bacilli  of  tuberculosis  in 
different  animals,  examined  under  the  same  conditions  of  amplifica- 
tion and  illumination,  x  1200.  Lamp-light  illumination. 

FIG.  1. — Bacilli  in  pus  from  the  wall  of  a  human  tubercular  cavity.  In 
this  specimen  the  bacilli  are  shorter  than  those  in  tubercular  sputum, 
and  are  very  markedly  beaded. 

FIG.  2. — Bacilli  in  pus  from  a  tubercular  cavity  from  another  case  in  man. 
They  are  present  in  the  preparation  in  enormous  numbers.  The  pioto- 
plasm  occupies  almost  the  whole  of  the  sheath,  and  the  bacilli  are 
strikingly  thin  and  long. 

FIG.  8.— Bacilli  in  sputum  from  an  advanced  case  of  phthisis,  showing 
the  ordinary  appearance  of  bacilli  in  sputum  ;  some  beaded,  others 
stained  in  their  entirety;  occurring  both  singly  and  in  pairs,  and 
in  groups  resembling  Chinese  letters. 

FIG.  4. — Bacilli  in  a  section  from  the  lung  in  a  case  of  tuberculosis  in  man. 
The  bacilli  in  human  tuberculosis  are  found  in,  and  between,  the  tissue 
cells  ;  and  sometimes,  as  in  equine  and  bovine  tuberculosis,  in  the 
interior  of  giant  cells,  but  not  so  commonly. 

FIG.  5. — From  a  cover-glass  preparation  of  the  deposit  in  a  sample  of  milk 
from  a  tubercular  cow.  The  bacilli  were  longer  than  the  average 
length  of  bacilli  in  bovine  tissue  sections,  and  many  were  markedly 
beaded. 


XXvi  DESCRIPTION  OF  PLATES. 

FIG.  (5. — From  a  section  of  the  brain  in  a  case  of  tubercular  meningitis  in  a 
calf,  showing  a  giant  cell  containing  bacilli  with  the  characters  usually- 
found  in  sections  of  bovine  tuberculosis. 

FIG.  7. — From  a  section  of  the  liver  of  a  pig  with  tubercle  bacilli  at  the 
margin  of  a  caseous  nodule. 

FIG.  8. — From  a  cover-glass  preparation  of  a  crushed  caseous  mesenteric 
gland  from  a  rabbit  infected  by  ingestion  of  milk  from  a  cow  with 
tuberculosis  of  the  udder. 

FIG.  9. — From  a  section  of  lung  in  a  case  of  equine  tuberculosis,  showing  a 
giant  cell  crowded  with  tubercle  bacilli. 

FIG.  10. — From  a  section  of  lung  from  a  case  of  tuberculosis  in  the  cat,  with 
very  numerous  tubercle  bacilli. 

FIG.  11. — From  a  cover-glass  preparation  of  a  crushed  caseous  nodule  from 
the  liver  of  a  fowl,  with  masses  of  bacilli.  These  are  for  the  most  part 
short,  straight  rods ;  but  other  forms,  varying  from  long  rods  to  mere 
granules,  are  also  found. 

FIG.  12. — From  sections  of  the  liver  and  of  the  lung  in  a  case  of  tubercu- 
losis of  a  Khea.  Isolated  bacilli  are  found,  as  well  as  bacilli  packed  in 
large  cells,  colonies  of  sinuous  bacilli,  and  very  long  forms  with  terminal 
spore-like  bodies  and  free  oval  grains. 

The  preparations  from  which  these  figures  were  drawn  were  all 
stained  by  the  Ziehl-Neelsen  method,  with  the  exception  of  the  first, 
which  was  stained  by  Ehrlich's  method. 


DESCRIPTION    OF    PLATE    XII. 

Tubercular  Mammitis. 

Following  p.  394. 

FIG.  1.— From  a  section  of  the  udder  of  a  milch  cow.  The  tubercular  deposit 
is  seen  to  invade  the  lobules  of  the  gland.  Lobules  comparatively  healthy 
are  marked  off,  more  or  less  sharply,  from  the  diseased  ones  in  which  the 
new  growth  in  its  progress  compresses  and  obliterates  the  alveoli.  Stained 
by  the  Ziehl-Neelsen  method  and  with  methylene-blue.  x  50. 

FIG.  2. — Part  of  the  same  preparation.  On  the  right  of  the  section  part  of  a 
healthy  lobule  is  seen.  On  the  left  a  lobule  is  invaded  by  tubercular  new 
growth  composed  of  round  cells,  epithelioid  cells  and  typical  giant  cells. 
Tubercle  bacilli  can  be  seen  both  singly  and  collected  in  groups.  They 
are  found  in  and  between  the  cells,  and  in  the  interior  of  giant  cells. 
Bacilli  may  be  seen  between  the  cells  lining  an  alveolus  and  projecting 
into  its  lumen,  x  800. 


DESCRIPTION    OF    PLATE    XIII. 

Tuberculosis  in  Swine. 

Following  p.  400. 

Section  of  liver  of  a  pig  with  scattered  tubercular  nodules.     Microscopical 
sections  of  the  liver  showed  tubercle  bacilli  in  very  small  numbers. 


DESCRIPTION   OF   PLATES.  XXvii 

DESCRIPTION    OF    PLATE    XIV. 
Bacillus  Leprae. 

Following  p.  408. 

FIG.  1. — From  a  section  of  the  skin  of  a  leper.  The  section  is,  almost  in 
its  entirety,  stained  red,  and,  with  moderate  amplification,  has  a  finely 
granular  appearance.  Stained  by  the  Ziehl-Neelsen  method  (carbolised 
fuchsine  and  methylene-blue).  x  200. 

FIG.  2. — Part  of  the  same  preparation  with  high  amplification,  showing  that 
the  appearances  described  above  are  due  entirely  to  an  invasion  of  the 
tissue  by  the  bacilli  of  leprosy,  x  1500. 

DESCRIPTION    OF    PLATES    XV.    AND    XVI. 

Actinomyces. 

Following  p.  432. 

PLATE  XV. 

FIG.  1.— From  a  preparation  of  the  grains  from  an  actinomycotic  abscess  in 
a  boy ;  examined  in  glycerine.  The  drawing  has  been  made  of  a  com- 
plete rosette  examined  by  focussing  successively  the  central  and  peripheral 
portions.  Towards  the  centre  the  extremities  of  the  clubs  are  alone 
visible ;  they  vary  in  size,  and  if  pressed  upon  by  the  cover-glass  give  the 
appearance  of  an  irregular  mosaic.  Towards  the  periphery  the  clubs  are 
seen  in  profile,  and  their  characteristic  form  recognised.  At  one  part 
there  are  several  elongated  elements,  composed  of  separate  links,  x  1200. 
FIG.  2. — Different  forms  of  clubs  from  preparations  in  which  the  rosettes  have 
been  flattened  out  by  gentle  pressure  on  the  cover-glass,  x  2500. 

(a)  Single  club.  (ft)  Bifid  club,  (c)  Club  giving  rise  to  four 
secondary  clubs.  (cT)  Four  clubs  connected  together,  recalling 
the  form  of  a  bunch  of  bananas.  (<?)  Mature  club  with  a  lateral 
bud.  (/)  Apparently  a  further  development  of  the  condition 
represented  at  (<?).  (^)  Club  with  a  lateral  bud  and  transverse 
segmentation,  (h)  Single  club  with  double  tranverse  segmenta- 
tion, (i)  Club  with  oblique  segmentation,  (j)  Collection  of 
four  clubs,  one  with  lateral  gemmation,  another  with  oblique 
segmentation.  (Ji)  Club  with  lateral  buds  on  both  sides,  and 
cut  off  square  at  the  extremity.  (I)  Club  with  a  daughter  club 
which  bears  at  its  extremity  two  still  smaller  clubs,  (ni)  Club 
divided  by  transverse  segmentation  into  four  distinct  elements. 
O)  Elongated  club  composed  of  several  distinct  elements.  (0)and 
(  j?)  Clubs  with  terminal  gemmation.  (#)  Palmate  group  of  clubs, 
(r)  Trilobed  club.  (*)  Club  with  apparently  a  central  channel. 
(£)  Filament  bearing  terminally  a  highly  refractive  oval  body. 

PLATE    XVI. 
FlG.  1. — From  a  section  of  a  portion  of  the  growth  removed  from  a  boy 

during  life.     The  tissue  was  hardened  in  alcohol,  and  cut  in   celloidin. 

The  section  was  stained  by  Gram's  method  and  with  orange-rubin.      x  50. 
FlG.  2. — From   the  same  section.     A  mass  of  extremely  fine  filaments  occupies 

the  central  part  of  the  rosette.     Many  of  the  filaments  have  a  terminal 

enlargement.     The  marginal  part  shows  a  palisade  of  clubs  stained  by  the 

orange-rubin.      x  500. 


XXV'iii  DESCRIPTION   OF   PLATES. 

FIGS.  3  and  4. — From  cover-glass  preparations  of  the  fungus  teased  out  of  the 
new  growths  produced  by  inoculation  of  a  calf  with  pus  from  a  boy 
suffering  from  pulmonary  actinomycosis.  Stained  by  Gram's  method  and 
orange-rubin.  The  threads  are  stained  blue  and  the  clubs  crimson  («) 
In  the  younger  clubs  the  thread  can  be  traced  into  the  interior  of  the 
club  (&).  In  some  of  the  older  clubs  the  central  portion  takes  a  yellowish 
stain,  and  in  others  the  protoplasm  is  not  continued  as  a  thread,  but  is 
collected  into  a  spherical  or  ovoid  or  pear-shaped  mass.  In  others,  again, 
irregular  grains  stained  blue  are  scattered  throughout  the  central  portion 
(Fig.  4).  x  1200. 

FIG,  5. — From  a  pure-culture  on  glycerine-agar.  (a)  branching  filaments, 
(ft)  a  mass  of  entangled  filaments.  Gram's  method,  x  1200. 

FIG.  (j. — From  a  similar  but  older  cultivation.  («)  a  filament  with  spores, 
(i)  chains  of  spores  simulating  streptococci.  Gram's  method,  x  1200. 


DESCRIPTION    OF    PLATES    XVII.    AND    XVIII. 

Actinomycosis  Bovis. 

Following  p.  434. 

PLATE   XVII. 

Section  of  an  actinomycotic  tongue  stained  by  the  method  of 
Gram  and  with  eosin. 

FIG.  1. — This  illustrates  the  appearance  which  is  usually  seen  under  a  low 
power,  when  a  section  is  stained  by  Gram's  method  and  with  eosin.  The 
central  portion  of  a  mass  of  the  fungus  is  either  unstained  or  tinged  with 
eosin,  while  the  marginal  portion  is  stained  blue.  The  reverse  is  seen,  as  a 
rule,  in  sections  from  man  ;  although  under  a  low  power  the  general  appear- 
ance of  sections  from  these  two  sources  is  somewhat  similar,  x  50. 

FIG.  2. — a,  b,  c,  d,  represent  the  earliest  recognisable  forms  of  the  ray  fungus 
in  the  interior  of  leucocytes.  In  e  the  club-forms  can  be  recognised.  In 
/  and  g  there  are  small  stellate  groups  of  clubs,  x  500. 

FIG.  3. — A  part  of  the  section  represented  in  Fig.  1,  under  a  high  power.  The 
marginal  line  of  blue  observed  under  a  low  power  is  now  recognised  as  the 
result  of  the  stain  being  limited  to  the  peripherally  arranged  clubs.  At 
(a)  part  of  a  rosette  has  undergone  calcification  ;  the  clubs  are  granular, 
and  have  not  retained  the  stain.  At  (V)  and  close  to  it  there  are  the 
remains  of  rosettes  in  which  the  process  of  calcification  is  almost  complete, 
x  500. 

PLATE   XVIII. 

The  figures  in  this  plate  are  taken  from  sections  of  a  case  of 
so-called  "  osteosarcoma,"  in  which  the  growth  of  the  fungus  was 
remarkably  luxuriant.  The  specimens  were  stained  by  Plauts' 
method. 

FIG.  1.— Different  forms  of  clubs  in  different  specimens  :    x  1200. 
(a)  Very  small  club-shaped  elements. 
(&)  A  club  with  transverse  segmentation. 
(<?)  A  club  with  lateral  daughter  clubs. 


DESCRIPTION    OF   PLATES.  XXIX 

(d  and  e)  Clubs  with  terminal  offshoots  resembling  teleutospores. 
(/)  A  club  with  developing  daughter  clubs  on  the  left,  and  on  the 

right  a  mature  secondary  club. 
(</)  A  segmental  club  with  lateral  offshoots. 
(/*)  Two  clubs  undergoing  calcification. 

FIG.  2.— A  very  remarkable  stellate  growth  comprised  of  nine  wedge-shaped 
collections  of  clubs  radiating  from  a  mass  of  finely  granular  material, 
x  500. 

FIG.  3. — A  rosette  undergoing  central  calcification;  and  consisting  in  part  of 
extremely  elongated  clubs  resembling  paraphyses.  Calcareous  matter  is 
also  being  deposited  in  the  club-shaped  structures,  x  500. 
FiG.  4. — Part  of  a  rosette  with  continuation  of  the  club-shaped  bodies 
into  transversely  segmented  branching  cells  apparently  representing  short 
hyphae.  x  500. 

FIG.  5. — A  rosette  from  another  section  in  which  similar  appearances  are 
observed  as  in  Fig.  4.  x  500. 

DESCRIPTION    OF    PLATE    XIX. 

Pure-cultivations  of  Actinomyces. 

Following  p.  488. 

These  tubes  were  selected  from  a  great  number  of  cultivations 
in  which  there  were  different  appearances.  In  some  instances  the 
growths  had  a  faint  tinge  of  pink. 

FIG.  1. — Pure-cultivation  on  the  surface  of  potato,  showing  a  luxuriant 
sulphur-yellow  growth  entirely  composed  of  entangled  masses  of  fila- 
ments. After  three  months'  growth. 

FIG.  2. — Pure-culture  from  the  same  series,  on  glycerine-agar.  In  this  case 
the  culture  remained  perfectly  white.  The  jelly  was  coloured  reddish- 
brown.  After  fifteen  months'  growth. 

FIG.  3.— Pure-culture  on  glycerine-agar  in  which  the  growth  was  dark- 
brown,  in  parts  black,  and  the  jelly  stained  dark-brown.  After  nearly 
two  years'  growth. 

DESCRIPTION    OF    PLATES    XX.    AND    XXI. 

Actinomycosis  Bovis. 

Following  p.  440. 

PLATE    XX. 

FIG.  1. — From  a  section  of  an  actinomycofeic  tongue  stained  by  the  triple 
method  (Ziehl-Neelsen,  logwood  and  orange-rubin).  In  this  section  the 
separate  centres  of  growth  are  clearly  shown.  Each  neoplasm  consists  of 
a  fungus  system,  in  which  the  masses  of  the  fungus,  situated  more  or  less 
centrally,  are  surrounded  with  round  cells,  epithelioid  cells,  sometimes 
giant  cells,  and  lastly  fibrous  tissue  forming  a  more  or  less  distinct 
capsule.  In  parts  the  fungi  have  fallen  out  of  the  section.  x  50. 

FIG.  2. — From  a  section  of  a  "tubercular"  nodule  from  the  lungs  of  a 
Norfolk  heifer  with  pulmonary  actinomycosis.  The  nodule  is  a  multiple 
growth  surrounding  a  bronchus,  and  is  enclosed  by  a  capsule,  in  the 


XXX  DESCRIPTION   OF   PLATES. 

vicinity  of  which  the  pulmonary  alveoli  are  compressed.  It  is  composed 
of  a  number  of  separate  neoplasms,  and  each  of  the  latter  is  composed  of 
secondary  centres  of  growth  resembling  the  giant-cell  systems  of  bacillary 
tuberculosis.  The  new  growth  is  composed  of  ray-fungi,  large  multi- 
nucleated  cells,  sometimes  .distinct  giant  cells,  round  cells,  epithelioid  cells, 
and,  surrounding  them,  fibrous  tissue.  On  examination  of  the  same 
specimen  with  a  higher  power  the  typical  rosettes  of  clubs  are  sometimes 
surrounded  by  multinucleated  cells,  and  sometimes  small  rosettes  are 
found  like  tubercle  bacilli,  in  the  interior  of  giant  cells.  From  a  pre- 
paration stained  by  Ziehl-Neelsen,  logwood,  and  orange-rubin.  x  50. 

PLATE  XXI. 

FiG.  1. — (a)  A  leucocyte  containing  the  fungus  in  its  earliest  recognisable 
form.  (J)  A  large  multinucleated  cell  containing  the  fungus  in  an  early 
stage  with  the  club-form  already  visible,  (c)  A  leucocyte  containing  a 
small  stellate  fungus,  (d)  A  large  cell  containing  clubs  arranged  in  a 
small  rosette,  (e)  A  multinucleated  cell  with  clubs  arranged  in  a  palmate 
form.  All  the  above  are  drawn  from  sections  of  actinomycotic  tongues 
stained  by  the  triple  method,  x  500. 

FIG.  2. — A  giant  cell  with  large  vesicular  nuclei  at  the  periphery,  and  in  the 
centre  a  fully  formed  rosette  of  actinomyces  with  a  smaller  growth  within 
a  "  daughter "  cell.  From  a  section  of  the  tongue  of  an  ox  stained  by 
the  triple  method,  x  500. 

FIG.  3. — A  very  large  circular  giant  cell,  with  its  ring  of  nuclei  at  the 
periphery,  enclosing  several  isolated  tufts  of  actinomyces.  From  a  section 
of  a  nodule  in  the  lung.  Stained  by  the  triple  method,  x  500. 

FIG.  4. — Three  rosettes  of  actinomyces  surrounded  by  a  row  of  large,  some- 
what angular  multinucleated  cells.  From  a  section  of  the  tongue  of  an 
ox  stained  by  the  triple  method,  x  430. 


DESCRIPTION    OF    PLATE    XXII. 

Bacillus  tetani. 

Following  p.  458. 

FIG.  L— From  a  cover-glass  preparation  of  a  pure-cultivation  of  the  tetanus 
bacillus  in  broth;  stained  with  Neelsen's.  carbolised  fuchsine.  x  1200. 
Lamplight  illumination. 

FIG.  2. — From  a  cover-glass  preparation  from  the  same  source  ;  stained  with 
Neelsen's  solution  and  methylene  blue,  x  1200.  Lamplight  illumination. 


PART     I. 

THEORETICAL   AND    TECHNICAL. 


OF  THF, 

UNIVERSITY 


BACTERIOLOGY 

AND 

INFECTIVE   DISEASES. 


CHAPTER  I. 

• 

HISTORICAL  INTRODUCTION. 

THE  researches  of  Pasteur  into  the  role  played  by  bacteria  in  the 
processes  of  fermentation  and  putrefaction,  and  the  investigations  of 
the  practical  mind  of  Lister,  with  the  resulting  evolution  of  antiseptic 
surgery,  demonstrated  the  necessity  for  a  more  intimate  acquaint- 
ance with  the  life-history  of  these  micro-organisms.  Further  re- 
searches in  diseases  such  as  anthrax,  the  silkworm  malady,  pyaemia, 
septicaemia,  and  fowl-cholera,  invested  the  science  of  Bacteriology 
with  universal  interest  and  vast  importance;  while  the  investiga- 
tions which  established  an  intimate  connection  between  bacteria 
and  other  infective  diseases,  and  more  especially  the  discovery  by 
Koch  of  bacteria  in  tuberculosis  and  in  Asiatic  cholera,  claimed  the 
attention  of  the  whole  thinking  world. 

Those  bacteria  which  are  connected  with  disease,  and  more 
especially  those  which  have  been  proved  to  be  the  causa  causans,  are 
of  predominant  interest  and  importance. 

The  first  attempt  to  demonstrate  the  existence  of  a  contagium 
vivum  dates  back  almost  to  the  discovery  of  the  microscope. 
Athanasius  Kircher,  nearly  two  and  a  half  centuries  ago,  expressed 
his  belief  that  there  were  definite  micro-organisms  to  which  diseases 
were  attributable.  The  microscope  had  revealed  that  all  decom- 
posing substances  swarmed  with  countless  micro-organisms  which 
were  invisible  to  the  naked  eye,  and  Kircher  sought  for  similar 
organisms  in  diseases  which  he  considered  might  be  due  to  their 
agency.  The  microscope  which  he  described  obviously  could  not 

1 


2  BACTERIOLOGY. 

admit  of  the  possibility  of  studying,  or  even  detecting,  the  micro- 
organisms which  are  now  known  to  be  associated  with  certain 
diseases  ;  and  it  is  not  surprising  that  his  teachings  did  not  at  the 
time  gain  much  attention.  They  were  destined,  however,  to  receive 
a  great  impetus  from  the  discoveries  which  emanated  from  "the 
father  of  microscopy." 

Antony  van  Leeuwenhoek  had  learned  as  a  youth  to  grind  and 
polish  lenses,  and  later  in  life  employed  his  spare  time  in  constructing 
microscopes,  and  in  conducting  those  researches  which  have  made 
for  him  a  name  which  is  familiar  to  all  microscopists.  His  researches 
were  published  in  a  series  of  letters  to  the  Royal  Society.  In  1675 
he  described  extremely  minute  organisms  in  rain-water,  well-water, 
infusions  of  pepper,  hay,  and  other  vegetable  and  animal  substances, 
in  saliva,  and  in  scrapings  from  the  teeth  ;  and,  further,  he  was 
able  to  differentiate  these  minute  living  things  by  their  size,  their 
form,  and  the  character  of  their  movements.  In  1683  these 
discoveries  were  illustrated  by  means  of  woodcuts,  and  there  can  be 
little  doubt,  from  the  drawings  of  these  micro-organisms,  that  they 
are  intended  to  represent  leptothrix  filaments,  vibrios,  and  spirilla. 
Indeed,  we  can  almost  recognise  these  micro-organisms  as  bacteria 
from  Leeuwenhoek's  graphic  descriptions,  apart  from  his  figures. 
They  were  described  as  moving  in  the  most  characteristic  manner, 
progressing  with  great  rapidity,  or  spinning  round  like  a  top,  and 
so  excessively  minute  that  they  were  only  perceived  with  great 
difficulty.  The  smallest  forms  could  hardly  be  examined  individually ; 
but,  viewed  en  masse,  they  closely  resembled  a  swarm  of  gnats  or 
flies.  In  another  communication,  published  in  1692,  he  gives 
some  idea  of  the  size  of  these  animalcules  by  stating  that  they 
were  a  thousand  times  smaller  than  a  grain  of  sand.  Others 
which  were,  comparatively  speaking,  of  considerable  length,  were 
characterised  by  their  peculiar  mode  of  progression,  bending  and 
rolling  on  themselves — movements  which,  he  adds,  created  both 
delight  and  astonishment  in  the  mind  of  the  observer.  Leeuwenhoek 
himself  was  not  disposed  to  believe  in  the  possibility  of  such 
organisms  being  found  in  the  blood  in  disease;  but  as  soon  as  he 
had  proved  the  actual  existence  of  such  minute  creatures,  theoretical 
physicians  were  not  wanting  who  at  once  attributed  various  maladies 
to  their  agency.  Among  these,  Nicholas  Andry  is  made  conspicuous 
by  his  work  published  in  1701.  Andry  classed  the  minute  organisms 
discovered  by  Leeuwenhoek  as  worms. 

In  1718  Lancisi  believed  that  the  deleterious  effect  of  the  air  of 
malarial  districts  depended  upon  animalcules,  and  others  considered 


HISTORICAL   INTRODUCTION.  $ 

that  the  plague  in  Toulon  and  Marseilles  in  1721  arose  from  a 
similar  cause.  In  fact,  by  some,  all  diseases  were  attributed  to- 
vermicules,  and  this  led  to  the  theory  being  ridiculed  and  discredited. 

In  spite  of  adverse  criticism,  the  theory  of  contagium  vivum 
survived,  and  Linnaeus  acknowledged  it  by  placing  the  micro- 
organisms discovered  by  Leeuwenhoek,  the  contagia  of  specific 
fevers,  and  the  causes  of  putrefaction  and  fermentation,  into  one 
class — "  chaos."  The  theory  was  further  supported  by  the  writings 
of  Plenciz,  who,  in  1762,  very  ably  discussed  the  nature  of  contagium,. 
as  well  as  the  relation  of  animalcules  to  putrefaction  and  disease. 
However,  no  proofs  in  support  of  these  theories  were  forthcoming, 
and  gradually  the  idea  of  contagium  vivum  fell  into  obscurity,  and 
indeed  came  to  be  regarded  by  some  as  an  absurd  hypothesis. 

Though  a  causal  relation  of  animalcules  to  diseases  was  for  a 
time  discredited,  the  natural  history  of  these  micro-organisms  was- 
studied  with  increasing  interest.  In  1778  Baron  Gleichen  described 
and  figured  a  great  number  of  micro-organisms  which  he  had 
discovered  in  various  vegetable  infusions.  Joblot,  Lesser,  Reaumur,. 
Hill,  and  many  others  worked  at  the  same  subject.  Hill  remarked 
that  there  was  hardly  the  least  portion  of  matter  or  the  least  drop 
of  fluid  of  any  kind  naturally  found  in  the  earth,  which  was  not 
inhabited  by  multitudes  of  animalcules.  But  these  observers  inclined 
rather  to  searching  for  new  forms  than  to  studying  more  thoroughly 
those  which  had  been  already  discovered ;  and,  as  a.  result,  but  little 
scientific  progress  was  made  until  the  time  of  Miiller,  of  Copen- 
hagen. Miiller,  in  1786,  criticised  the  work  of  previous  writers, 
and  pointed  out  that  they  had  been  too  much  occupied  with  merely 
finding  new  micro-organisms.  Miiller  took  into  account  the  form 
of  the  micro-organism,  its  mode  of  progression,  and  other  biological 
characters,  and  on  such  data  based  a  classification.  Thus  the 
scientific  knowledge  of  these  minute  beings  was  considerably  advanced 
by  his  writings  and  illustrations. 

The  subject  which  now  eclipsed  all  others  in  interest  was  the 
origin  of  these  micro-organisms.  Two  rival  theories  were  widely 
discussed — spontaneous  generation,  and  development  from  pre-exist- 
ing germs ;  and  the  researches  that  were  made  in  the  course  of 
this  discussion,  and  the  discoveries  which  resulted,  indirectly  yet 
materially  advanced  the  germ  theory  of  disease,  and  explain  many 
of  the  phenomena  in  the  life-history  of  the  pathogenic  microbes 
which  have  been  brought  to  light  in  recent  years. 

Spontaneous  development  of  micro-organisms  in  putrescible 
infusions  was  believed  in  by  many,  but  was  supported  by  no  one 


4  BACTERIOLOGY. 

with  greater  persistency  than  Needham.  Needham  found  that 
animalcules  readily  developed  when  meat  infusion  was  boiled  an 
transferred  to  a  well-stoppered  flask,  and  he  could  only  explain  this 
by  supposing  that  they  originated  spontaneously  from  the  material  of 
the  infusion.  In  1768  Bonnet  strenuously  opposed  these  conclusions 
on  purely  theoretical  grounds,  and  maintained  that  it  was  far  more 
probable  that  the  ova  of  the  animalcules  were  present  in  the  infusions 
or  were  suspended  in  the  air  enclosed  in  the  flask. 

Spallanzani  was  the  first  to  demonstrate  by  experiment  the 
correctness  of  Bonnet's  arguments.  It  occurred  to  him  to  boil  the 
infusion  in  flasks,  and  to  seal  the  vessels  during  tJie  process  of  boiling. 
As  a  result  the  flasks  remained  free  from  putrefaction,  and 
animalcules  only  developed  when  the  infusion  was  exposed  to  the  air 
by  making  a  hole  in  the  flask.  That  Spallanzani's  experiments 
were  reliable,  and  his  conclusions  correct,  was  evidenced  by  the  fact 
that  his  simple  precaution  led  to  great  practical  results,  as  Fra^ois 
Appert  introduced,  on  this  principle,  the  method  of  preserving  meats, 
vegetables,  and  other  provisions. 

The  disciples  of  Needham  nevertheless  brought  forward  counter 
objections.  Treviranus  urged  that  a  certain  quantity  and  quality  of 
air  was  necessary  for  the  spontaneous  development  of  these  infusoria, 
and  that  by  sealing  the  flasks,  too  small  a  quantity  of  air  was  in 
contact  with  the  infusion,  and,  further,  that  this  air  had  become 
•changed  in  quality  by  the  process  of  boiling. 

Spallanzani  argued  against  these  objections,  but  did  not  support 
his  opinions  by  further  experiments,  so  that  the  question  remained 
for  a  time  undecided. 

In  1836  Francis  Schulze  devised  an  experiment  which  brought 
still  further  evidence  against  Neeclham's  theory.  Schulze  filled  a 
glass  vessel  half  full  with  distilled  water  and  different  animal  and 
vegetable  substances.  This  was  plugged  with  a  doubly-bored  cork, 
and  through  each  perforation  a  glass  tube  was  introduced,  bent  at 
a  right  angle.  On  boiling  the  flask,  steam  issued  freely  from  each 
tube,  and  all  parts  were  thoroughly  sterilised.  Each  tube  was  then 
connected  with  a  bulbed  tube,  one  bulb  containing  concentrated 
sulphuric  acid  and  the  other  a  solution  of  potash.  Fresh  air  was 
drawn  into  the  flask  by  aspiration,  and  this  was  deprived  of  any 
germs  which  might  be  present  by  its  passage  through  the  sulphuric 
acid.  The  result  was  that  the  infusion  .remained  without  any 
development  of  micro-organisms.  When,  on  the  other  hand,  air  was 
admitted  without  first  being  drawn  through  the  sulphuric  acid,  the 
n  usion  in  a  short  time  teemed  with  animalcules.  In  other  words, 


HISTORICAL   INTRODUCTION.  O 

Schulze  demonstrated  that  in  spite  of  free  access  to  air,  ivhich  Jiad  not 
been  heated,  the  infusions  remained  free  from  germs. 

Schwann,  in  1837,  arrived  at  similar  results.  He  found  that 
putrescible  substances  remained  sterile  if  exposed  to  an  abundant 
supply  of  air  which  was  heated  by  being  passed  through  a  melted  mix- 
ture of  metals.  This  convinced  him  that  the  cause  of  the  decompo- 
sition which  would  otherwise  have  occurred  must  exist  in  the  air. 

The  objection  remained  that  in  the  experiments  of  Schulze  and 
Schwann,  the  air  which  was  admitted  to  the  flasks  had  undergone 
either  a  chemical  or  a  thermal  change,  and  therefore  the  theory  of 
Needham  was  not  yet  entirely  disposed  of. 

In  1854  the  final  blow  was  dealt  by  Schroder  and  Van  Dusch, 
These  investigators  demonstrated  that  decomposition  could  be  obviated 
without  resorting  either  to  thermal  or  chemical  treatment  of  the 
air,  as  simple  filtration  of  the  air  through  cotton-wool  was  shown  to- 
be  efficacious  in  excluding  germs.  Finally,  Hoffman  in  1860,  and 
independently,  Chevreuil  and  Pasteur  in  1861,  showed  that  even 
cotton-wool  could  be  dispensed  with,  as  a  sterile  solution  would 
remain  sterile  when  the  neck  of  the  vessel  was  bent  into  an 
S -shaped  curve.  Micro-organisms  in  the  air  entering  the  flask 
were  deposited  by  gravitation  in  the  bend  of  the  tube. 

The  advocates  of  spontaneous  generation  were  ready  with  fresh 
objections.  They  now  urged  that  the  medium  lost  its  power  of 
undergoing  decomposition  by  being  boiled.  This  objection  was  at 
once  set  aside  by  the  fact  that  when  unfiltered  air  was  admitted  to- 
the  infusion,  decomposition  set  in.  Additional  evidence  was  brought 
against  spontaneous  generation  by  the  experiments  of  Pasteur, 
Burdon  Sanderson,  Lister,  and  others,  in  which  it  was  shown  that 
blood,  urine,  and  milk  would  remain  without  decomposition,  when  all 
precautions  were  adopted  to  avoid  contamination  in  filling  the 
sterilised  flasks. 

Even  at  this  stage  of  this  great  scientific  controversy  fresh 
difficulties  arose,  for  it  was  found  that  in  certain  solutions  which  had 
been  boiled  and  hermetically  sealed  in  flasks  micro-organisms  made 
their  appearance.  In  1872  Charlton  Bastian  published  a  research 
which  was  to  prove  that  spontaneous  generation  actually  took 
place.  Decoctions  of  turnip  and  cheese  which  had  been  filtered, 
neutralised,  and  boiled  for  ten  minutes,  and  hermetically  sealed 
during  the  boiling,  were  found  after  a  time  to  contain  micro 
organisms.  These  results,  however,  were  before  long  explained  by 
the  fact  that  in  milk,  infusions  of  hay,  and  certain  other  decoctions, 
the  spores  of  bacilli  are  present,  which  are  much  more  resistant 


6  BACTERIOLOGY. 

than  the  bacilli  themselves.  In  such  cases  mere  scalding  or  boiling 
for  a  few  minutes  will  not  sterilise  the  solution.  The  bacilli  are 
destroyed,  but  not  their  spores;  and  if  the  latter  remain  unhurt, 
they  will  germinate,  and  rapidly  multiply.  But  if,  as  Tyndall 
found,  the  boiling  be  repeated  a  second  and  a  third  time,  all  the 
spores  will  be  destroyed ;  for  in  the  intervals  between  the  boilings 
the  spores  sprout  into  bacilli,  and  the  bacilli  at  the  next  boiling 
perish  ;  so  that  after  three  or  four  repeated  boilings  the  infusion  is 
rendered  perfectly  free  from  germs. 

While  this  discussion  was  occupying  the  attention  of  the  whole 
scientific  world,  some  investigators  had  been  again  following  up  the 
theory  of  a  connection  between  micro-organisms  and  disease. 

In  1837  Oagniard  Latour  and  Schwann  independently  made  the 
•discovery  that  the  yeast  plant  was  a  living  organism,  and  the  true 
cause  of  yeast  fermentation.  The  close  analogy  between  the  pro- 
cesses of  fermentation  and  of  certain  diseases  had  long  been  held ; 
and,  therefore,  when  it  was  proved  that  fermentation  was  due  to  a 
micro-organism,  fresh  advocates  appeared  in  support  of  the  theory 
that  diseases  were  produced  by  similar  agencies.  Boehm,  in  1838, 
•described  certain  organisms  in  cholera,  which  was  at  that  time 
raging  in  Europe ;  but  the  researches  of  Bassi,  who  a  year 
previously  had  discovered  the  cause  of  a  disease  of  silkworms, 
attracted  much  greater  attention. 

Bassi  discovered  that  in  this  disease  extremely  minute  spores 
existed  on  the  bodies  of  the  worms,  which  were  conveyed  from  the 
sick  to  the  healthy.  They  destroyed  the  healthy  worms  by 
germinating  in  their  skins  and  growing  into  their  bodies.  These 
discoveries  may  be  said  to  have  brought  the  theory  of  contagium 
vivum  to  life  again  ;  and  Henle,  in  reviewing  the  facts  of  the  case  in 
1 840,  came  to  the  conclusion  that  the  cause  of  all  contagious  diseases 
must  be  of  a  living  nature,  and  this  he  maintained,  although  he 
had  searched  in  vaccine  and  small-pox  lymph,  in  the  desquamation 
of  scarlet  fever,  and  in  other  diseases  without  success. 

Bassi's  discovery  and  Henle's  doctrine  encouraged  a  number  of 
investigators,  and  remarkable  results  followed.  In  favus,  in  herpes 
tonsurans,  in  pityriasis  versicolor,  fungus  threads  and  spores  were 
found,  and  were  regarded  as  being  of  etiological  importance, 
inasmuch  as  the  morbid  lesions  corresponded  with  the  growth  of  the 
particular  fungus. 

Cholera  became  especially  a  subject  for  research.  Swaine, 
Brittan,  and  Budd  found  micro-organisms  in  choleraic  dejecta. 
Davaine  described  certain  monads  in  the  intestinal  contents,  but  no 


HISTORICAL    INTRODUCTION.  7 

causal  connection  was  established  between  these  organisms  and  the 
disease  ;  and  when  the  cholera  disappeared  the  interest  in  contagium 
rim  HI  \vaiied,  and  was  eclipsed  by  the  question  of  fermentation. 
The  discoveries  which  followed  in  this  subject  had  a  very  important 
1) :«aring  on  the  micro-parasitic  origin  of  communicable  diseases. 

Pasteur,  following  up  the  researches  of  Cagniard  Latour  and 
Schwann,  demonstrated  in  1857  that  the  lactic,  acetic,  and  butyric 
fermentations  were  produced  by  micro-organisms. 

Previously  to  this,  in  1850,  Davaiiie  and  Rayer  had  noted  the 
existence  of  little  rod-like  or  filamentous  bodies  about  the  size  of  a 
blood  corpuscle  in  the  blood  of  a  sheep  that  had  cjiecl  of  splenic  fever. 
Pollender  had  seen  similar  bodies  in  the  blood  of  cows.  Davaine 
did  not  at  first  pay  much  heed  to  this  discovery;  but  in  1863  he 
thoroughly  reinvestigated  the  subject,  and  conducted  a  series  of 
experiments  which  led  him  to  the  conclusion  that  the  actual  cause 
of  splenic  fever  was  an  organised  being  whose  presence  and 
multiplication  in  the  blood  produced  changes  in  that  fluid  of  the 
nature  of  fermentation,  resulting  in  the  death  of  the  animal. 

These  conclusions  were  not  accepted  by  all,  and  indeed,  the 
evidence  was  so  far  incomplete  that  sceptics  were  justified  in  con- 
sidering that  these  experiments  afforded  only  a  working  hypothesis. 
But  Davaine's  comparison  between  this  disease  and  fermentation 
attracted  the  attention  of  Pasteur,  whose  mind  had  been  fully  trained 
for  entering  upon  this  investigation  by  the  researches  which  he  had 
been  carrying  on  in  the  interval  between  Davaine's  publications  of 
1857  and  1863. 

Pasteur,  as  already  mentioned,  had  been  working  at  fermentation, 
and  his  attention  was  next  directed  to  studying  the  so-called  diseases 
of  wines,  and  subsequently  to  a  contagious  disease  which  committed 
ravages  among  silkworms.  By  laborious  researches  Pasteur  was 
able  to  confirm  the  belief  that  this  disease  of  silkworms  was  due  to 
the  presence  of  micro-organisms  discernible  with  the  aid  of  the  micro- 
scope. These  oval  shining  bodies  in  the  moth,  worm,  and  eggs  had 
been  previously  observed  by  Cornalia,  and  described  by  Nageli  as 
Nosema  bombycis,  and  by  Lebert  as  Panhistophyton.  But  it  was 
reserved  for  Pasteur  to  introduce  a  means  of  combating  the  disease. 
Pasteur  showed  that  when  a  silkworm,  whose  body  contained  these 
micro-organisms,  was  pounded  up  with  water  in  a  mortar,  and  the 
mixture  painted  with  a  brush  on  the  leaves  on  which  healthy  wrorms 
were  fed,  they  would  all  without  fail  succumb  to  the  disease. 

As  the  contagious  particles  were  transmitted  to  the  eggs,  a 
method  for  preventing  the  spread  of  the  disease  suggested  itself. 


8  BACTERIOLOGY. 

Each  female  moth  was  kept  separate  from  the  others,  and  allowed  to 
deposit  her  eggs  on  a  small  linen  cloth.  The  moth  was  then  pinned 
to  the  corner  of  the  cloth,  and  left  for  future  examination.  When 
the  time  for  this  arrived,  the  moth  was  crushed  up  with  water  in  a 
mortar,  and  a  drop  examined  under  the  microscope.  When  any 
trace  of  corpuscular  matter  was  found  to  be  present,  the  cloth  with 
its  collection  of  eggs  was  burnt ;  and  if  not,  the  eggs  were  set  aside 
for  use. 

Complete  as  this  appears  to  be  as  a  demonstration  of  a  causal 
connection  between  the  micro-organisms  and  the  disease,  it  could 
obviously  be  objected  that  there  was  no  distinct  proof  that  the 
corpuscular  bodies  constituted  the  actual  contagium.  There  was  no 
isolation  of  the  organisms,  no  artificial  cultivation  of  them  apart 
from  the  diseased  moth  or  worm,  and  subsequent  production  of  the 
disease  by  means  of  the  isolated  organisms.  The  same  objection 
was  applicable  to  Davaine's  investigations.  Davaine  found  rods 
in  association  with  anthrax,  and  maintained  that  they  were 
causally  related  ;  but  others  stated  that  it  was  possible  to  inoculate 
animals  with  anthrax  blood  containing  rods,  and  to  produce  the 
disease  without  being  able  to  detect  the  rods  again  in  the  blood 
of  the  animal  experimented  upon.  It  was  also  urged  that  it  was 
possible  to  infect  with  anthrax  blood  after  the  rods  had  disappeared, 
and  to  find  a  reappearance  of  the  bacilli  in  the  blood  of  the 
inoculated  animal. 

The  well-known  fact  that  anthrax  was  especially  prevalent  in 
certain  seasons  and  certain  localities  appeared  to  lend  great  support 
to  these  objections.  The  disease,  in  fact,  was  regarded  by  some 
as  originating  from  peculiar  conditions  of  climate  and  soil.  The 
fallacies  in  these  objections  were,  however,  rapidly  dispelled. 
Bellinger,  in  1872,  pointed  out  that  the  blood,  from  which  the  rods 
had  disappeared,  was  still  virulent  owing  to  the  presence  of  the 
spores  of  the  bacillus,  and  that  it  was  owing  to  the  soil  being  impreg- 
nated with  these  spores  that  the  disease  broke  out  in  certain 
localities.  Yet  there  still  remained  many  who  refused  to  regard 
these  particles  as  living  bodies,  some  looking  upon  them  simply  as 
crystals ;  and  the  question  of  their  importance  remained  undecided 
for  several  years. 

In  1877  Robert  Koch  published  a  memoir  in  which  he  fully 
described  the  life-history  of  the  anthrax  or  splenic  fever  bacillus, 
and  gave  a  complete  demonstration  of  the  life-history  of  the  micro- 
organism, and  the  definite  proofs  of  its  pathogenic  properties.  He 
pointed  out  how  the  rods  grew  in  the  blood  and  tissues  by  lengthen- 


HISTORICAL   INTRODUCTION.  9 

ing  and  by  cross  division.  Further,  that  in  the  blood  or  in  serum 
or  in  aqueous  humour  they  not  only  grew  into  long  leptothrix 
filaments,  but  they  produced  enormous  numbers  of  seeds  or  spores. 
He  traced,  by  continuous  observation  on  the  warm  stage,  the  whole 
life  cycle,  from  the  fission  of  the  rods  to  the  formation  of  spores  and 
the  sprouting  of  the  spores  into  fresh  rods.  Further,  he  carried 
011  the  disease  by  inoculating  from  mouse  to  mouse  for  several 
generations,  and  observed  that  in  the  blood  of  the  animal  and  in 
the  swollen  spleen  the  glass-like  rods  were  always  to  be  found. 

Pasteur  also  studied  the  microbe  of  splenic  fever,  and  amply 
confirmed  and  extended  the  observations  of  Koch  by  his  researches 
on  the  attenuation  of  the  anthrax  virus. 

Pasteur  also  met  with  adverse  criticism.  Paul  Bert  argued 
that  the  bacilli  were  of  no  importance,  because  he  could  destroy 
them  by  exposing  material  containing  them  to  great  pressure,  and 
yet  the  material  produced  the  disease  on  inoculation.  But  such 
measures  did  not  destroy  the  spores;  and  finally,  Paul  Bert  was 
convinced  of  his  error  when  Pasteur  demonstrated  cultures  of  the 
anthrax  bacillus  in  urine,  from  which  successive  generations  were 
started,  and  that  with  such  cultivations  the  disease  could  always  be 
produced. 

It  was,  however,  principally  the  researches  of  Koch  which 
placed  the  doctrine  of  contagium  vivum  on  a  scientific  basis. 

Koch's  improvements  in  the  methods  of  cultivation,  his  recom- 
mendation of  the  necessary  microscopical  apparatus,  his  histological 
methods  for  examining  these  minute  organisms,  and  his  famous 
postulates  for  proving  beyond  controversy  the  existence  of  specific 
pathogenic  micro-organisms,  elevated  the  theory  of  contagium  vivum 
to  a  demonstrated  and  established  fact.  The  chain  of  evidence 
regarded  by  Koch  as  essential  for  proving  the  existence  of  a 
pathogenic  organism  was  as  follows  :— 

1.  The   micro-organism  must  be  found  in  the  blood,  lymph,  or 
diseased  tissue  of   man  or  animal  suffering  from    or  dead   of   the 
disease. 

2.  The  micro-organisms  must  be  isolated  from  the  blood,  lymph, 
or  tissues,  and  cultivated  in  suitable  media — i.e.,  outside  the  animal 
body.    These  pure  cultivations  must  be  carried  on  through  successive 
generations  of  the  organism. 

3.  A  pure  cultivation  thus  obtained  must,  when  introduced  into 
the  body  of  a  health)-  animal,  produce  the  disease  in  question. 

4.  In    the    inoculated   animal   the   same   micro-organism    must 
again  he  found. 


10  BACTERIOLOGY. 

The  chain  of  evidence  is  still  more  complete  if  we  can  from 
Artificial  cultures  obtain  a  chemical  substance  which  is  capable 
of  producing  the  disease  independently  of  living  micro-organisms. 

It  is  of  very  little  value  merely  to  detect  or  artificially  to 
cultivate  a  bacterium  associated  with  disease.  We  must  endeavour 
to  establish  the  exact  relationship  of  the  bacteria  to  disease  processes, 
and  the  determination  of  the  true  pathogenic  microbe  is  beset 
with  fallacies.  In  many  diseases  bacteria  have  been  regarded  as 
the  actual  contagia,  until  a  searching  inquiry  by  other  investigators 
has  shown  that  the  evidence  was  most  unsatisfactory  or  entirely 
misleading.  For  example,  in  diseases  with  lesions  of  the  external 
or  internal  linings  of  the  body,  extraneous  micro-organisms  may 
get  into  the  circulation  and  be  swept  into  the  internal  organs, 
where  they  either  perish  in  the  battle  with  the  healthy  tissues 
which  are  opposed  to  their  existence,  or  they  may  gain  the  upper 
hand,  and  set  up  destructive  processes.  Such  organisms,  when 
found  in  association  with  these  diseases,  may  be  discovered  in  the 
blood  and  internal  organs  ;  and  though  only  accidental  epiphytes, 
often  associated  with  septic  complication,  they  may  too  readily  be 
accepted  by  the  enthusiast  as  the  actual  contagium  of  the  disease 
in  question. 

It  is  only  when  such  fallacies  are  exposed  that  we  are  brought 
once  more  face  to  face  with  the  fact  that  the  nature  of  the  contagium 
in  hydrophobia,  variola,  vaccinia,  scarlet  fever,  measles,  and  many 
other  diseases,  is  still  undetermined. 


CHAPTER   II. 

MORPHOLOGY    AND   PHYSIOLOGY    OF   BACTERIA. 

BACTERIA  may  be  considered  as  minute  vegetable  cells  destitute  of 
nuclei.  They  are  distinguished  from  animal  cells  by  being  able  to 
<lerive  their  nitrogen  from  ammonia  compounds,  and  they  differ 
from  the  higher  vegetable  cells  in  being  unable  to  split  up  carbonic 
acid  into  its  elements,  owing  to  the  absence  of  chlorophyll.  Yon 
Engelmann  and  Van  Tieghem  include  among  the  bacteria  certain 
organisms,  named  by  them  Bacterium  chlorimim,  Bacterium  viride, 
and  Bacillus  virens,  which  are  coloured  green  by  this  substance,  but 
it  is  quite  possible  that  they  may  be  Algae,  and  further  researches 
are  required  before  any  conclusions  are  definitely  arrived  at  as  to 
the  exact  place  these  particular  organisms  occupy  in  the  vegetable 
kingdom. 

Composition. — For  our  knowledge  of  the  chemical  composition 
•of  bacteria  we  are  chiefly  indebted  to  Nencki.  Their  constituents 
are  found  on  analysis  to  vary  slightly,  according  to  whether  the 
bacteria  are  in  zoolgoea  or  in  the  active  state.  In  the  latter  condition 
they  are  said  to  consist  of  83*42  per  cent,  of  water.  In  one  hundred 
parts  of  the  dried  constituents  there  are  the  following  :— 

A  nitrogenous  body    .         .         .         .         .  84*20 

Fat     .         . 6-04 

Ash 4-72 

Undetermined  substances     ....  5'04 

This  nitrogenous  body  is  called  myco-protein,  and  consists  of 

Carbon 52'32 

Hydrogen    .         .  .         .         .         .  7*55 

Nitrogen      .         .         .         .         .         .         .         14' 75 

but  no  sulphur  or  phosphorus. 

The  nitrogenous  body  appears  to  vary  in  different  species,  for 
in  Bacillus  anthracis  a  substance  has  been  obtained  which  does  not 

11 


12  BACTERIOLOGY. 

give  the  reactions  of  myco-protein,  and,  therefore,  is  distinguished 
as  anthrax-protein. 

Considering  bacteria  as  cells,  we  may  speak  of  the  cell-wall  and 
the  cell-contents.  The  cell-wall  consists  of  cellulose,  or,  according 
to  Nencki,  in  the  putrefactive  bacteria  of  myco-protein.  It  may  be 
demonstrated  by  the  action  of  iodine,  which  contracts  the  proto- 
plasmic contents,  and  renders  the  cell- wall  visible.  By  staining 
cover-glass  preparations  of  the  anthrax  bacillus  by  the  method  of 
Gram,  the  rods  are  at  first  uniformly  stained,  by  subjecting  them  to 
iodine  solution  the  protoplasmic  contents  are  contracted,  and  alcohol 
decolorises  the  sheath,  which  may  be  then  stained  in  contrast,  with 
eosin. 

The  cell- wall  may  be  either  pliable  or  rigid.  Pliability  is 
observed  in  the  long  filaments,  which  are  endowed  with  a  slow 
vermicular  movement,  while  rigidity  accounts  for  the  maintenance 
of  the  characteristic  form  of  several  species,  such  as  spirilla. 

The  cell-protoplasm  yields  inyco- protein.  In  some  it  is  homogene- 
ous, and  in  others  granular.  The  action  of  the  aniline  dyes  indicates 
a  close  relation  to  nuclear  protoplasm,  though  all  nuclear  stains  are 
not  suitable  for  bacteria.  In  some  cases  also  the  bacteria  remain 
stained  under  the  influence  of  a  reagent,  which  removes  the  colour 
from  nuclei.  The  power  of  fixing  the  stain  is  not  always  present, 
and  indicates  a  difference  in  the  protoplasm  of  different  species. 
Thus  in  staining  phthisical  sputum,  the  nitric  acid  removes  the 
stain  from  all  bacteria  and  bacilli  present,  with  the  exception  of  the 
tubercle  bacillus.  This  difference  in  the  protoplasm  of  different 
species  is  also  illustrated  by  the  necessity,  in  many  cases,  of  using 
special  processes,  owing  to  the  ordinary  methods  being  unsatisfactory 
or  not  producing  any  result. 

The  protoplasm  of  some  bacteria  contains  starch  granules  ;  thus 
Clostridium  butyricum  gives  the  starch  reaction  with  iodine. 
Sulphur  granules  are  present  in  some  species  of  Beggiatoa  which 
thrive  in  sulphur  springs.  The  colouring-matter  of  the  pigmented 
bacteria  is  probably  external  to  the  cell  as  a  rule  :  for  example,  in 
Micrococcus  prodigiosus  the  pigment  granules  are  distictly  between 
the  cells ;  on  the  other  hand,  in  Beggiatoa  roseo-persicina,  or  the 
peach-coloured  bacterium,  the  special  pigment  bacterio-purpurin 
appears  to  be  dissolved  in  the  cell  protoplasm.  In  Bacillus 
pyocyaneus  the  pigment  is  certainly  not  localised  entirely  in  the 
cell,  for  it  becomes  rapidly  diffused  in  the  surrounding  medium, 
considerably  beyond  the  confines  of  the  growth  itself. 

In  several  species,  either  as  a  result  of  a  secretion  from  the  cell  or 


MORPHOLOGY   AND   PHYSIOLOGY   OF   BACTERIA.  13 

of  the  absorption  of  moisture  and  consequent  swelling  of  the  outer 
layer  *of  the  cell- wall,  a  mucinous  or  gelatinous  envelope  develops 
around  them.  This  envelope  may  form  a  capsule,  such  as  we  meet 
with  in  certain  bacteria  found  in  the  rusty  sputum  of  pneumonia,  and 
in  Micrococcus  tetragenus  ;  or  it  may  occur  as  a  continuous  sheath 
around  a  chain  of  bacteria,  which  by  its  disappearance  sets  free 
the  individual  links.  The  capsule  is  soluble  in  water,  and  under 
some  circumstances  is  difficult  to  demonstrate.  In  the  pneumo- 
coccus  of  Friedlander  the  capsule  disappears  on  cultivation,  but 
reappears  in  preparations  made  from  an  inoculated  animal.  In  the 
pleuritic  fluid  of  a  mouse  these  cocci  are  often  found  with  a  parti- 
cularly well-marked  capsule,  and  in  other  encapsuled  cocci  the  extent 
of  the  envelope  has  been  observed  to  vary  considerably  in  the  same 
species  of  bacterium. 

When  this  gelatinous  material  forms  a  matrix,  in  which 
numbers  of  bacteria  are  congregated  in  an  irregular  mass,  we  have 
what  is  termed  a  zoogloea.  The  zooglcean  stage  is  a  resting  stage, 
often  preceded  or  followed  by  a  motile  stage.  Thus  bacteria  may 
be  present  in  a  solution  in  an  active  state,  and  after  a  time  a  scum 
or  pellicle  forms  on  the  surface  of  the  liquid,  which  consists  of 
zooglcea.  At  the  edges  of  the  zoogloea,  individuals  may  be  seen  to 
again  become  motile,  and  after  detaching  themselves  to  swim  off  in 
the  surrounding  fluid. 

The  zoogkean  stage  may  be  observed  sometimes  in  cultivations  in 
broth,  and  also  in  nutrient  gelatine  which  has  become  liquefied, 
The  inoculated  bacteria  grow  and  multiply,  and  after  a*  time  a  film 
appears  on  the  surface  of  the  liquefied  layer.  In  cultivations  on 
potato  the  appearances  in  this  stage  are  varied,  and  sometimes 
extremely  characteristic.  In  the  case  of  a  bacillus  which  readily 
develops  on  unsterilised  potatoes,  the  zooglcea  may  spread  over  the 
cut  surface,  forming  a  pellicle  which  can  be  raised  en  masse  like 
a  delicate  veil.  Another  bacillus  forms  a  zoogloea,  consisting  of  a 
tenacious  layer  which  can  be  drawn  out  in  long  stringy  threads. 
In  Ascococcus  Billrothii  the  gelatinous  envelope  develops  to  such 
an  enormous  extent  that  it  forms  the  characteristic  feature  of  the 
species.  (Fig.  1.)  . 

Form. — The  individual  cells  vary  in  form,  and  may  either 
remain  isolated  or  attached  to  each  other.  Round  cells  and  egg- 
shaped  cells  are  called  cocci.  The  spherical  form  is  the  most 
common,  but  cocci  are  occasionally  exclusively  ovoid,  as  in  Strepto- 
coccus bombycis.  The  giant  cocci  of  some  species  are  spoken  of  as 
megacocci,  to  distinguish  them  from  the  ordinary  cocci,  or  micrococci. 


OF   THR 

TTNTVF"RQTrPVr 


14 


BACTERIOLOGY. 


The  fission  by  which  the  cocci  increase  may  take  place  in  one 
direction  only,  and  if  the  two  resulting  cells  remain  attached  to 
each  other  they  form  a  diplococcus.  If  these  two  cells  again  divide, 
and  the  resulting  cells  remain  linked  together,  we  get  a  chain  or 
rosary,  termed  streptococcus.  These  chains  may  consist  of  a  few 
individuals  linked  together,  or  of  several  hundreds,  in  which  case 
the  chains  are  generally  curved  or  twisted.  When  the  division 
occurs  in  two  directions,  so  that  four  cocci  result,  a  tetrad  or 
merismopedia  is  formed  ;  when  in  three  directions,  one  coccus  divides 
into  eight,  and  the  result  is  a  packet  form  or  sarcinacoccus. 
Immediately  after  division,  the  daughter  cells  are  not  perfectly 
circular,  but  are  flattened  or  facetted  where  they  are  opposite  to 


FIG.  1.— Ascococcus  BILLROTHII,    x   65.     [After  Cohn.] 


each  other.  They  gradually  become  rounded  off,  and  each  daughter 
cell  is  then  ready  to  divide  in  its  turn.  In  other  cases  the  cocci 
after  division  only  form  irregular  heaps  or  collections  like  bunches 
of  grapes.  This  form  is  sometimes  distinguished  as  staphylococcus, 
but  it  cannot  be  considered  an  important  feature.  When  we  find 
irregular  masses  of  cocci  united  by  intercellular  substance  and 
embedded  in  a  tough  gelatinous  matrix,  the  form  is  described  as 
ascococcus. 

Another  type  is  the  rod,  characteristic  of  bacterium  and  bacillus. 
The  rods  may  vary  considerably  in  length.  The  very  short  rods 
with  rounded  ends  are  difficult  to  distinguish  from  the  oval  cocci, 
but  differ  in  that  a  rod,  however  short  it  may  be,  must  have 
two  sides  parallel.  The  vibrio  or  bent  rod  may  be  considered  as 
the  connecting  link  between  the  rods  and  the  corkscrew  forms  or 


DESCRIPTION    OF    PLATE    I. 
Bacteria,  Schizomycetes,  or  Fission  Fungi. 

1.  Cocci  singly  and  varying  in  size.  2.  Cocci  in  chains  or  rosaries  (strepto- 
coccus). 3.  Cocci  in  a  mass  (staphylococcus).  4  and  5.  Cocci  in  pairs 
(diplococcus).  6.  Cocci  in  groups  of  four  (merismopedia).  7.  Cocci  in  packets 
(sarcina).  8.  Bacterium  termo.  9.  Bacterium  termo  x  4000  (Dallinger  and 
Drysdale).  10.  Bacterium  septiccemice  hccmorrliagicce.  11.  Bacterium  pneu- 
iiumice  crouposcB.  12.  Bacillus  subtilis.  13.  Bacillus  murisepticus.  14. 
Bacillus  diphtheria.  15.  Bacillus  typliosus  (Eberth).  16.  Spirillum  undula 
(Cohn).  17.  Spirillum  volutans  (Cohn).  18.  Spirillum  cliolerce  Asiaticce. 
19.  Spirillum  Obermeieri  (Koch).  20.  Spirocliceta  plicatilis  (Fliigge).  21. 
Vibrio  rugula  (Prazmowski).  22.  Cladothrix  Forsteri  (Cohn).  23.  Cladothrix 
dichotoma  (Cohn).  24.  Nonas  Okenii  (Cohn).  25.  Nonas  Warmingii  (Cohn). 
26.  Rhabdomonas  rosea  (Cohn).  27.  Spore-formation  (Bacillus  alvei).  28. 
Spore -formation  (Bacillus  anthracis).  29.  Spore-formation  in  bacilli  cultivated 
from  a  rotten  melon  (Frankel  and  Pfeiifer).  ,30.  Spore-formation  in  bacilli 
cultivated  from  earth  (Frankel  and  Pfeiffer).  31.  Involution-form  of  Crenotlirix 
(Zopf).  32.  Involution-forms  of  Vibrio  serpens  (Warming).  33.  Involution- 
forms  of  Vibrio  rugula  (Warming).  34.  Involution-forms  of  Clostridium 
polymyxa  (after  Prazmowski).  35.  Involution-forms  of  Spirillum  cholerce 
Asiaticce.  36.  Involution-forms  of  Bacterium  aceti  (Zopf  and  Hansen). 
37.  Spirulina-form  of  Beggiatoa  alba  (Zopf).  38.  Various  thread-forms  of 
Bacterium  merismopedioides  (Zopf).  39.  False-branching  of  Cladothrix  (Zopf). 


Plate  I. 


MAC'TF.KIA,  SCHI7OMYCETES,  OH  FISSION    FUNGI. 


MORPHOLOGY   AND   PHYSIOLOGY   OF   BACTERIA.  15 

Lastly,  we  have  the  filamentous  forms,  which  may  be 
straight,  leptothrix,  or  wavy,  spirochceta,  or  the  wavy  thread  may 
be  looped  and  entwined  on  itself,  spirulina. 

The  term  involution  / orm  is  applied  to  certain  peculiar  shapesr 
which  result  more  especially  in  bacteria  grown  under  abnormal 
conditions.  They  are  round,  oval,  pear-shaped,  or  club-formed 
enlargements. 

Movement. — Many  bacteria  are  devoid  of  movement  through- 
out the  whole  of  their  life  history.  Others,  during  certain  stages  of 
their  life  cycle,  and  possibly  some  forms  always,  are  endowed  with 
locomotive  power.  The  character  of  the  movement  is  very  varied,, 
and  ranges  from  a  slow  undulatory  motion  to  one  of  extreme 
rapidity.  Many  appear  to  progress  in  a  definite  direction.  Others 
move  continuously,  first  in  one  direction  and  then  in  another,  and 
others  again  seem  to  hesitate  before  altering  their  course.  They 
may  either  glide  along  smoothly  or  progress  with  a  tremulous  motion. 


FIG.  2.— SPIROCH.ETA  FROM  SEWAGE  WATER,   x   1200. 

They  appear  to  be  able  to  avoid  obstacles,  and  to  set  themselves 
free  from  objects  with  which  .they  have  accidentally  come  into 
contact.  Vibrios  have  a  peculiar  serpentine  movement,  but  other 
forms,  such  as  the  commonly  known  Bacterium  termo  and  segments 
of  spirilla,  such  as  comma -bacilli,  revolve  around  their  long  axis 
as  well  as  make  distinct  progression.  The  complete  spirilla  are 
characterised  by  the] familiar  corkscrew  movement.  With  regard 
to  cocci  there  is  some  doubt  as  to  whether  they  are  endowed  with 
independent  movement,  any  quivering  or  oscillation  being  generally 
regarded  as  only  Brownian  or  molecular.  In  some  straight  thread- 
forms,  which  are  motile,  the  movement  is  very  slow  and  vermicular 
in  character,  but  in  wavy  threads,  such  as  the  Spirochseta  plicatilisr 
there  is  not  only  an  undulatory  motion,  with  rapid  progression  across 
the  field  of  the  microscope,  but  if  they  are  confined  by  more  or 
less  debris,  they  give  very  peculiar  and  characteristic  spasmodic 
movements.  (Fig.  2.) 

The  rod-forms  of  Proteus  vulgaris  exhibit  very  extraordinary 


16 


BACTERIOLOGY. 


movements  on  the  surface  of  solid  nutrient  gelatine.  Groups  of 
rods  may  be  observed  to  pass  each  other  in  opposite  directions. 
Single  individuals  meet  and  progress  side  by  side,  or  one  or  more 
individuals  may  part^from  a  group  and  glide  away  independently. 
Occasionally  a  number  of  rods  progress  in  single  file.  It  is,  however, 
difficult  to  believe  that  these  movements  can  occur  on  a  solid  surface. 


FIG.  3. — FLAGELLA. 

1.  Coccus  with  flagellum.  2.  Similar  coccus  dividing,  with  two  flagella.  3.  Colony 
of  flagellated  macrococci  of  Begyiatoa  roseo-persicina.  4.  Short  rod  from  the 
same  Beggiatoa  with  flagella  [all  after  Zopf].  5.  Bacillus  with  flagella  [from 
a  photograph  by  Koch}.  6.  Bacillus  subtilis  [after  Brefeld].  7,  8.  Short  rod- 
forms  of  Beggiatoa  roseo-persicina  with  one  flagellum  [after  Zopf].  9.  Very 
long  rod  of  the  same,  with  flagellum  at  both  ends  [after  Warming].  10.  Vibrio, 
with  double  flagellum  at  each  end  [after  Warming].  11.  Vibrio,  with  flagella 
[from  a  photograph  by  the  author].  12.  Spirillum  with  flagella  [from  a  photo- 
graph by  Koch].  13.  Spirillum  with  flagella  [after  Zopf].  14.  Spirillum  with 
double  flagella  [after  Zopf].  15.  Beggiatoa  roseo-persicina,  with  a  triple 
flagellum  at  one  end  ;  and  16,  with  a  double  flagellum  at  both  ends  [after 
Warming]. 

The  author  is  inclined  to  believe  that  there  is  an  almost  inappreciable 
layer  of  liquid  on  the  surface  of  the  gelatine,  which  is  expressed 
after  the  gelatine  sets.  In  tubes  of  nutrient  agar-agar  gelatinised 
obliquely  and  then  kept  upright  the  liquid  so  expressed,  collects  at 
the  bottom  of  the  sloping  surface. 


MORPHOLOGY    AND    PHYSIOLOGY    OF   BACTERIA. 


17 


The  means  by  which  bacteria  are  endowed  with  the  power  of 
spontaneous  movement  and  of  progression  may  still  be  said,  in 
some  cases,  to  be  unsettled.  The  author  has  watched  the  move- 
ment of  long  slender  threads  in  sewage- contaminated  water,  which 
could  only  be  explained  by  the  inherent  contractility  of  the  proto- 
plasmic contents  ;  for  if  any  drawing  or  propelling  organ  existed 
in  proportion  to  the  length  of  the  organism,  it  would  probably  have 
been  visible.  But  in  many  cases  the  organism  is  provided  with  a 
vibratile  lash  or  flagellum  at  one  end,  or  with  one  or  more  at  both 
ends,  or  with  numerous  lateral  and  terminal  flagella. 

Some  observers  believe  that  the  movement  of  cocci  is  due  to  the 


FIG.  4. — BACILLUS  MEGATHERIUM. 

a.  A  chain  of  rods,  x  250.     The  rest  x  600. 

b.  Two  active  rods. 

d  to/.  Successive  stages  of  spore-formation. 
h  to  ni.  Successive  stages  of  germination. 

[After  De  Bary.] 

existence  of  a  flagellum.  In  Bacterium  termo  the  existence  of  a 
lash  at  either  end  was  first  determined  by  the  researches  of  Dallinger 
and  Drysdale.  In  motile  bacilli,  such  as  the  hay  bacillus  and 
Bacillus  ulna,  and  in  vibrios  and  spirilla,  the  flagella  can  be  readily 
recognised  by  expert  microscopists  with  the  employment  of  the  best 
lenses,  and,  what  is  of  equal  importance,  proper  illumination.  They 
are  objects  of  extreme  delicacy  and  tenuity,  and  in  stained  prepara- 
tions may  be  absent  from  retraction  or  injury.  Koch  succeeded 
in  photographing  them  after  staining  with  logwood,  which  turned 
them  a  brown  colour.  The  author  has  observed  them  in  vibrios  in 
preparations  stained  with  gentian  violet,  from  which  also  they  have 
been  photographed,  in  spite  of  the  violet  colour,  by  the  use  of 

2 


18 


BACTERIOLOGY. 


isochromatic  dry  plates,  and  more  recently  special  methods  have 
been  introduced,  by  Loftier  arid  others,  by  which  they  can  be  stained 
and  photographed  with  comparative  facility. 

It  is  not  certain  whether  the  flagella  are  extensions  of  the  cell- 
wall,  or  derived  from  the  internal  protoplasm.  Van  Tieghem  holds 
the  first  view,  and  does  not  regard  them  as  motile  organs  at  all. 
Zopf,  on  the  other  hand,  adheres  to  the  second  view,  and  moreover 
believes  that  they  can  be  retracted  within  the  cell-wall. 

Reproduction. — Bacteria  multiply  by  fission  and  by  processes 
which  may  be  considered  as  representing  fructification.  The 


FIG.  5. — CLOSTRIDIUM  BUTYRICUM,    x   1020. 

B.  Stages  of  spore-formation. 

C.  Stages  of  germination. 

;[ After  Prazmowski.] 


bacteria  exhibiting  the  latter  processes  have  been  divided  into  two 
groups,  distinguished  by  the  formation  of  endospores  in  the  one  and 
of  arthrospores  in  the  other.  In  the  process  of  fission  the  cell  first 
increases  in  size,  and  a  transverse  septum  forms  from  the  cell- wall, 
dividing  the  internal  protoplasm  into  two  equal  parts  ;  these  may 
separate  and  lead  an  independent  existence,  or  remain  linked 
together.  In  chains  of  cocci  the  individual  cells  are  easily  visible 
and  distinct,  but  in  the  thread- forms  resulting  from  the  linking 
together  of  rods,  as  in  the  anthrax  bacillus,  the  composition  of  the 
thread  is  only  demonstrated  by  the  action  of  reagents. 

Endospore  formation    may  be  conveniently  studied  in  Bacillus 
anthracis,  Bacillus  megatherium,  or  Bacillus  subtilis.      The.proto- 


MORPHOLOGY    AND    PHYSIOLOGY   OF   BACTERIA.  19 

plasm  becomes  granular,  and  at  certain  points  in  the  thread  a  speck 
appears,  which  gradually  enlarges  and  develops  into  a  circular 
or  egg-shaped,  sharply  denned,  highly  refractive  body.  The  spore 
grows  at  the  expense  of  the  protoplasm  of  the  cell,  which  in  time, 
together  with  the  cell-wall,  entirely  disappears,  and  the  spore  is  set 
free.  These  phenomena  are  best  seen  in  an  immotile  bacillus  in  a 
drop- cultivation  on  a  warm  stage ;  the  whole  process  may  then  be 
observed  continuously  from  beginning  to  end.  Spores  may  form  in 
each  link  of  the  thread,  so  that  a  regular  row  results,  or  they  may 
occur  at  irregular  intervals.  Spore-formation  also  occurs  in  bacilli 
which  do  not  develop  into  leptothrix  filaments.  The  spores  may 
develop  in  the  centre  or  at  one  end  of  the  rod.  In  the  tetanus 
bacillus  a  spore  develops  at  the  extreme  end,  producing  the  appear- 
ance of  a  drum-stick.  The  spore  may  be  considerably  wider,  but 
is  never  longer  than  the  parent  cell. 


FIG.  6. — LEUCONOSTOC  MESENTEROIDES  ;  COCCI-CHAINS  WITH  ARTHROSPORES 
(after  Van  Tieghem  and  Cienkowski). 

Arthrospore  formation  is  illustrated  in  Leuconostoc  mesenteroides. 
•Certain  elements  in  the  chain  of  cocci,  apparently  not  differing  from 
the  rest,  become  larger,  with  tougher  walls,  and  more  refractive. 
The  remaining  cells  die,  and  these  cells  having  acquired  the  pro- 
perties of  spores  are  set  free,  and  can  reproduce  a  new  growth  in 
any  fresh  nourishing  soil.  That  this  occurs  in  all  species  which 
do  not  form  endospores  is  at  present  only  a  supposition. 

Spores  are  invested  by  a  thick  membrane,  which  is  believed  to 
consist  of  two  layers.  To  this  they  probably  owe  the  property  they 
possess  of  retaining  vitality  when  desiccated,  and  of  offering  a 
greater  resistance  to  the  action  of  chemical  reagents  and  heat  than 
the  parent  cells. 

Spore -formation  has  been  regarded  by  some  as  occurring  when 
the  nourishing  soil  is  exhausted,  thus  providing  for  the  perpetuation 


20  BACTERIOLOGY. 

of  the  species.  In  anthrax  the  bacilli  do  not  form  spores  in  the 
living  body,  but  when  the  animal  dies  the  development  of  spores 
takes  place,  and  hence  the  danger  of  contaminating  the  soil  if  the 
body  is  disposed  of  by  burial.  Klein,  however,  has  pointed  out  that 
if  mice  and  guinea-pigs  which  have  died  of  anthrax  are  kept  un- 
opened, the  bacilli  simply  degenerate  and  ultimately  disappear. 
Thus  there  is  good  reason  for  believing  that  spore-formation  is 
not  due  to  exhaustion  of  the  pabulum,  but  probably  free  access  to 
oxygen  constitutes  an  important  factor  in  inducing  this  condition. 
If  we  inoculate  a  potato  with  anthrax,  copious  spore- formation 
occurs,  though  we  cannot  consider  that  the  nourishing  soil  has  been 
exhausted.  But  we  have  in  this  case  the  surface  of  the  potato 
freely  exposed  to  the  air  in  the  damp  chamber.  In  the  same  way, 
in  cultivations  on  agar-agar  solidified  obliquely  (so  as  to  get  a  large- 
surface),  spore-formation  readily  takes  place.  Contamination  of 


FIG.  7. — SPORE-BEARING  THREADS  OF  BACILLUS  ANTHRACIS,  DOUBLE- STAINED 
WITH  FUCHSINE  AND  METHYLENE  BLUE,  X  1200. 

the  ground  results,  therefore,  from  animals  in  which  a  post-morteni 
examination  has  been  made  and  the  blood  and  organs  freely  exposed 
to  the  air  ;  or  from  carcasses  the  hides  of  which  have  been  soiled 
with  excretions,  and  with  blood  which  issues  from  the  mouth  and 
nostrils  before  death. 

When  spores  are  introduced  into  a  suitable  medium  at  a  favour- 
able temperature  they  develop  again  into  rods.  The  spore  loses  its 
sharp  contour,  and,  at  one  pole  or  on  one  side,  a  pale  process  bursts 
through  the  membrane,  gradually  growing  into  a  rod  from  which 
the  empty  capsule  is  thrown  off. 

Spores  differ  from  the  parent  cells  in  their  behaviour  to  staining 
reagents.  Like  them,  they  can  be  stained  with  aniline  dyes,  but 
not  by  the  ordinary  processes.  They  require  to  be  specially  treated. 
This  is  probably  due  to  the  tough  capsule,  which  must  be  altered 
or  softened  by  heat  or  strong  acid,  until  it  allows  the  stain  to 
penetrate. 


MORPHOLOGY    AND   PHYSIOLOGY   OF   BACTERIA.  21 

Once  stained,  they  again  differ  from  the  parent  cells  in  resisting 
decolonisation  ;  this  fact  is  taken  advantage  of  to  double-stain  spore- 
bearing  bacilli  (Fig.  7). 

In  staining  micro-organisms,  the  protoplasm  is  sometimes  broken 
up  into  irregular  segments  or  granules,  as  in  many  spirilla,  and  we 
may  add  the  bacilli  of  tuberculosis  and  leprosy.  The  beaded 
appearance  of  the  tubercle  bacillus  is  well  known.  Some  observers 
have  regarded  the  beads,  others  the  bright  spaces  between  them, 
as  spores.  But  spores  in  unstained  preparations  appear  as  glistening 
bodies  with  sharp  contour.  They  do  not  stain  at  all,  or  very  little, 
by  the  ordinary  processes.  These  considerations  led  the  author  to 
stain  and  examine  tubercular  sputum  and  pure-cultures  under 
careful  illumination,  and  with  such  lenses  as  Powell  and  Lealand's 
2*g-  in.  horn.  imm.  The  tubercle  bacillus  in  sputum  (Fig.  8),  as  a  rule, 


FIG.  8.— BACILLI  OF  TUBERCLE  IN  SPUTUM,    x   2500  (from  photographs). 

consists  of  a  very  delicate  sheath,  holding  together  a  number  of 
deeply  stained  granules,  for  the  most  part  round  or  cylindrical,  with 
irregular  contour,  and  differing  considerably  in  size,  while  the  light 
interspaces  are  seen  to  vary  in  form  according  to  the  shape  of  the 
granules.  On  the  other  hand,  particularly  in  old  cultures,  more  or 
less  spherical,  sharply  denned  bodies  are  observed  in  the  bacilli,  and 
also  set  free.  These  are  the  true  spores  of  the  tubercle  bacillus, 
and  are  quite  distinct  from  the  irregular  granules.  There  can  be 
no  doubt  that  a  tubercle  bacillus  consists  of  a  very  delicate  sheath, 
with  protoplasmic  contents  which  have  a  great  tendency  to  break 
up  or  coagulate  into  little  segments  or  roundish  granules,  partly 
owing  to  their  age  and  the  conditions  under  which  they  are  grown, 
and  partly  to  the  treatment  they  are  subjected  to  in  making  a 
microscopical  preparation.  This  does  not  always  occur,  for  the 
bacilli  at  times  are  not  beaded,  but  are  stained  in  their  entirety. 
In  the  leprosy  bacilli  a  similar  appearance  occurs.  In  stained 


22  BACTERIOLOGY. 

sections  the  rods  have  a  beaded  appearance,  but  the  intervals  between 
the  granules  are  sometimes  very  long,  and  occasionally  the  protoplasm 
appears  to  have  collected  only  at  the  extreme  ends  of  the  rod. 

The  appearances  in  the  case  of  the  bacillus  of  glanders  and  the 
bacillus  of  hsemorrhagic  septicaemia  may  be  similarly  explained. 

The  fact  that  tubercular  sputum  preserves  its  virulence  for 
several  months,  even  after  desiccation,  is  to  be  attributed  to  the 
formation  of  spores.  Babes  claims  to  have  succeeded  in  differen- 
tiating them  by  double  staining. 

In  his  definition  of  spirilla,  Zopf  gives  the  spore-formation  as- 
absent  or  unknown.  In  comma-bacilli  in  sewage  water  the  author 
has  often  noted  appearances  suggestive  of  refractive  spores ;  and 
the  same  also  may  be  observed  in  vibrios,  differing  by  their  regular 
contour  from  the  irregular  spaces  occasionally  observed  in  stained 
preparations ;  but  they  are  only  vacuoles. 


FIG.  9.— COMMA-BACILLI  IN  SEW-  FIG.  10.  VIBRIOS  IN  WATER  CON- 

AGE    WATER,   STAINED    WITH  TAMINATED     WITH    SEWAGE, 

GENTIAN  VIOLET,    x   1200.  x  1200. 

Respiration  and  Nutrition. — Like  all  a-chlorophyllous  vegeta- 
bles, bacteria  require  for  their  nutrition  oxygen,  nitrogen,  carbon, 
water,  and  certain  mineral  salts.  Many  require  free  access  to  oxygen, 
others  can  derive  it  from  the  oxidised  compounds  in  the  medium 
in  which  they  grow.  Pasteur  divided  bacteria  into  two  great  classes 
— the  aerobic  and  anaerobic,  and  considered  that  the  latter  net 
only  had  no  need  of  oxygen,  but  that  its  presence  was  actually 
deleterious.  Though  this  view  must  be  considerably  modified,  the 
terms  are  convenient,  and  are  still  retained.  They  are  well  illus- 
trated by  the  bacillus  of  anthrax,  and  the  bacillus  of  malignant 
oedema;  and  a  simple  plan  of  demonstration  has  been  employed 
by  the  author.  A  fragment  of  tissue  from  the  spleen,  for  example, 
known  to  contain  anthrax  bacilli,  is  deposited  with  a  sterilised 
inoculating  needle,  with  the  necessary  precautions,  on  the  surface  of 
nutrient  agar-agar  in  a  test-tube ;  another  tube  of  nutrient  agar- 
agar  is  liquefied,  and  when  cooled  down  almost  to  the  point  of 


MORPHOLOGY   AND   PHYSIOLOGY    OF   BACTERIA.  23 

gelatinisation,  a  part  is  poured  into  the  first  tube,  so  that  when  it 
sets  the  piece  of  tissue  is  completely  embedded.  A  piece  of  tissue 
from  an  animal  suffering  from  malignant  oedema  is  treated  in  the 
same  way,  and  the  tubes  are  placed  in  the  incubator.  If  we 
examine  them  after  two  or  three  days,  we  shall  find  no  change  in 
the  anthrax  tube ;  the  bacillus  being  eminently  aerobic,  no  growth 
whatever  has  occurred.  In  the  tube  containing  the  bacilli  of 
malignant  cedema  there  will  be  a  more  or  less  characteristic 
cultivation. 

The  nitrogen  which  is  essential  for  building  up  their  protoplasm 
can  be  obtained  either  from  albumins,  or  from  ammonia  and  its 
derivatives.  That  the  albumins  can  be  dispensed  with  was  shown 
by  Pasteur,  who  employed  an  artificial  nourishing  solution  consti- 
tuted upon  a  formula  representing  the  essential  food  constituents. 

Carbon  is  derived  from  such  substances  as  cane  sugar,  milk 
sugar,  and  glycerine,  and,  in  some  cases,  by  the  splitting  up  of 
complex  proteid  bodies. 

Water  is  essential  for  their  growth,  but  deprivation  of  water 
does  not  kill  all  bacteria.  Desiccation  on  potato  is  employed  for 
preserving  some  micro-organisms,  as  a  new  growth  can  be  started, 
when  required,  by  transferring  some  of  the  dried  potato  to  fresh 
nourishing  ground.  Comma-bacilli,  on  the  other  hand,  are 
destroyed  by  drying.  Sugar  is  used  in  making  preserves,  because 
by  abstracting  water  it  prevents  the  development  of  micro- 
organisms. 

Mineral  or  inorganic  substances,  such  as  compounds  of  sodium 
and  potassium,  and  different  phosphates  and  sulphates,  are  necessary 
in  small  proportions. 

CIRCUMSTANCES  AFFECTING  THE  GROWTH  OF  BACTERIA. 

Xature  of  the  Soil. — Though  we  know  the  elements  necessary, 
we  are,  nevertheless,  as  yet  unable  to  provide  a  pabulum  suitable  for 
all  kinds  of  bacteria.  Thus  we  are  quite  unable  to  cultivate  some 
species  artificially.  Others  will  only  grow  upon  special  media. 
Many  grow  upon  nutrient  gelatine ;  but  some  species  only  if  it  be 
acid  or  alkaline  respectively.  Whether  in  the  latter  case  this  is  due 
purely  to  the  reaction  or  to  the  presence  of  the  particular  ingredients 
is  an  unsettled  point.  Though  the  comma-bacillus  of  Koch,  like  the 
majority  of  organisms,  grows  best  on  an  alkaline  medium,  yet  it 
is  \\ell  known  to  flourish  at  the  temperature  of  the  blood  on  the 
surface  of  potato,  which  is  acid. 


24  BACTERIOLOGY. 

Temperature. — The  influence  of  temperature  on  bacteria  will  be 
found  to  vary  according  to  the  species,  but  still  for  the  majority  we 
may  distinguish  a  maximum,  optimum,  and  minimum  temperature. 

Many  grow  best  at  the  temperature  of  the  blood,  and  hence  the 
value  of  nutrient  agar-agar,  which  is  not  liquefied  at  37°  C.  The 
tubercle  bacillus  will  only  grow  satisfactorily  at  a  temperature 
varying  between  30°  C.  and  41°  C.  On  the  other  hand,  many  forms 
grow  between  the  limits  of  5°  C.  and  45°  C.  At  these  temperatures 
their  functional  activity  is  paralysed,  but  they  are  not  destroyed, 
for  by  removal  to  favourable  conditions  they  spring  again  into  life. 
Bacteria  seem  to  have  -a  special  power  of  resisting  the  effects  of  cold. 
It  has  been  stated  that  comma-bacilli  exposed  to  a  temperature 
of— 10°  C.  for  an  hour,  and  bacilli  of  anthrax  after  exposure  to  a 
temperature  of  — 110°  C.,  still  retained  their  vitality.  Temperatures 
over  50°  to  60°  C.  destroy  most  bacteria,  but  not  their  spores  ;  spores 
,of  anthrax  retain  their  vitality  after  immersion  in  boiling  water,  but 
are  destroyed  by  prolonged  boiling.  Roughly  speaking,  all  patho- 
genic bacteria  grow  best  at  the  temperature  of  the  blood,  and 
non-pathogenic  bacteria  at  the  ordinary  temperature  of  the  room. 

Movement. — Bacteria  probably  grow  best  when  left  undisturbed. 
Violent  agitation  of  a  vessel  in  which  they  are  growing  certainly 
retards  their  growth,  but  a  steady  movement  is  stated  not  to  affect 
it ;  at  any  rate,  anthrax  bacilli  grow  with  enormous  rapidity  in  the 
blood-vessels,  in  spite  of  the  circulation. 

Compressed  Air. — Paul  Bert  maintained  that  a  pressure  of 
twenty-three  to  twenty-four  atmospheres  stopped  all  development 
of  putrefactive  bacteria.  Oxygen,  under  a  pressaire  of  five  or  six 
atmospheres,  is  stated  to  stop  their  growth.  Other  observers  have, 
however,  obtained  different  results. 

Gases. — Hydrogen  and  carbonic  acid  are  stated  to  stop  the 
movements  of  the  motile  bacteria.  Chloroform  is  believed  to  arrest 
the  changes  brought  about  by  the  zymogenic  species. 

Electricity. — Cohn  and  Mendelssohn  found  that  a  constant 
galvanic  current  produced  a  deleterious  effect  owing  to  electrolysis. 
At  the  positive  pole  the  liquid  became  distinctly  acid,  and  at  the 
negative  pole  distinctly  alkaline.  With  a  weak  current  there 
appeared  to  be  no  effect,  two  powerful  cells  at  the  very  least  being 
necessary. 

Light. — Downes  has  shown  that  sunlight  is  fatal  to  putrefactive 
bacteria.  This  is  believed  to  be  due  to  a  process  of  induced  hyper- 
oxidation,  from  which  living  organisms  ordinarily  are  shielded  by 
protective  developments  of  the  cell-wall,  or  of  colouring-matter, 


MORPHOLOGY   AND   PHYSIOLOGY   OF   BACTERIA.  25 

which  cat  off  injurious  rays.  Duclaux  has  investigated  the  same 
subject,  and  observed  that  micrococci  were  more  sensitive  to  sun- 
light than  the  spore-bearing  bacilli.  Engelmann  has  described  a 
bacterium  whose  movements  cease  in  the  dark,  and  Zopf  states  that 
in  his  cultures  of  Beggiatoa  roseo-persicina  the  growth  was  much 
more  strongly  developed  on  the  side  of  the  vessel  facing  the  light. 
Arloing,  Marshall  Ward,  and  Dieudonne  have  studied  the  effect 
of  the  sun's  rays  on  anthrax  spores,  and  on  chromogenic  and 
other  bacteria,  and  maintain  that  they  are  bactericidal.  The 
effect  is  due  chiefly,  if  not  entirely,  to  the  blue  rays. 

Chemical  Reagents. — Many  substances,  such  as  carbolic  acid, 
corrosive  sublimate,  chlorine,  bromine,  have  a  marked  effect  upon 
the  growth  of  bacteria.  This  will  be  more  fully  described  in. 
another  chapter.  In  several  cases  the  bacteria  themselves  secrete 
a  substance  which  is  injurious  to  their  future  development. 

PRODUCTS  OF  GROWTH. 

Bacteria  may  be  grouped  together  according  to  the  changes  pro- 
duced in  the  media  in  which  they  grow.  Thus  we  have  pigment- 
forming,  phosphorescent,  fermentative,  putrefactive,  nitrifying,  and 
disease-producing  bacteria. 

Chromogenic  or  pigment -forming  bacteria  elaborate  during  their 
growth  definite  colour  stuffs.  Such  species  are  exemplified  by  Bacillus 
violaceus,  which  produces  a  striking  purple  growth ;  Bacillus 
pyocyaneus,  which  secretes  pyocyanin,  a  substance  which  has  been 
isolated  and  obtained  in  a  crystalline  form  ;  Micrococcus  prodigiosus, 
which  produces  a  pigment  allied  to  fuchsine  ;  Beggiatoa  roseo-per- 
sicina, which  is  characterised  by  the  presence  of  bacterio-purpurin ; 
Sarcina  lutea,  Bacillus  cyanogenus,  and  many  others. 

Photogenic,  or  light-producing,  bacteria  are  found  more  especially 
in  sea -water.  There  are  several  species  of  phosphorescent  bacilli, 
and  according  to  Beyrinck  the  best  medium  for  their  cultivation  is 
fish-broth  made  with  sea-water.  Photographs  can  be  obtained  of 
cultures  by  their  own  light. 

Zymogenic  or  ferment  bacteria  produce  their  changes  in  non- 
nitrogenised  media.  Bacterium  aceti,  by  its  growth  produces  the 
acetic  fermentation  in  wine  by  which  alcohol  taking  up  atmospheric 
oxygen  is  converted  into  vinegar  : — 

C2H60    +   O2  =  C2H402  +   H20. 

The  fermentation  of  urine,  by  which  urea  is  converted  into  carbonate 
of  ammonia,  can  be  brought  about  by  several  micro -organisms,  but 


26  BACTERIOLOGY. 

notably  by  the  Bacterium  urese.  The  change  produced  is  represented 
by  the  following  formula  :  — 

CO{NIP  *  2H2°  =  (NH<)  2C°3- 

Clostridium  butyricum  converts  the  salts  of  lactic  acid  into- 
butyric  acid,  producing  the  butyric  fermentation  in  solutions  of 
starch,  dextrine,  and  sugar.  These  bacteria  are  agents  in  the 
ripening  of  cheese,  and  the  production  of  sauerkraut.  Thus,  in  a 
solution  neutralised  with  calcium  carbonate  :  — 

2[Ca(C3H503)2]  +  H20  =  Ca(C4H7O-)2  +  CaCO3  +  3C02  +  H8. 

In  the  so-called  viscous  fermentation  of  wines,  Streptococcus  viscosus- 
produces  a  gummy  substance.  According  to  Pasteur,  the  change 
may  be  thus  represented  :  — 

25(C12H22011)  +  25(H20)  =  12(C12H20010)  +  24(C6H1406)  + 
12(C02)  +  12(H2O). 

And  as  another  example,  the  Bacillus  acidi  lactici  may  be  mentioned  f 
through  the  agency  of  which  sugar  of  milk  is  converted  into  lactic 
acid  :  — 


Saprogenic  or  putrefactive  bacteria  play  a  most  important  part 
in  the  economy  of  nature.  They  produce  changes  allied  to  fermenta- 
tion in  complex  organic  substances.  Their  action  on  proteids, 
according  to  Hoppe-Seyler,  may  be  compared  to  digestion  ;  bodies 
like  peptones  are  first  produced,  then  leucin,  tyrosin,  and  fatty 
acids  ;  lastly  indol,  phenol,  sulphuretted  hydrogen,  ammonia,  carbonic 
acid,  and  water.  They  abstract  the  elements  they  require,  and  the 
remainder  enter  into  new  combinations.  Associated  with  the  forma  - 
tion  of  these  substances  are  certain  bodies  which  have  a  poisonous 
effect  when  introduced  into  animals.  These  poisonous  alkaloids,. 
ptomaines,  produce  a  septic  poisoning,  which  must  be  distinguished 
from  septic  infection.  The  effects  of  septic  poisoning  depend  on  the 
dose,  whereas  the  effects  of  septic  infection  are,  to  a  certain  extent, 
independent  of  the  dose.  A  small  quantity  of  a  septic  poison  may 
produce  only  transient  effects,  and  a  relatively  large  quantity  may 
be  necessary  to  produce  vomiting,  rigors,  arid  death.  Septic  in- 
fection, on  the  other  hand,  may  result  equally  from  a  small  doser 
because  the  poison  introduced  is  a  living  organism  which  is  capable 
of  propagation  and  multiplication.  Our  knowledge  of  these 
alkaloids  is  largely  attributable  to  the  researches  of  Selmi,  Gautier, 
and  Brieger,  and  the  result  of  their  work  will  be  referred  to  again. 


MORPHOLOGY   AND   PHYSIOLOGY    OF   BACTERIA.  27 

Nitrifying  bacteria  play  a  very  important  part  by  providing 
plant  life  with  a  most  necessary  food.  They  occur  in  the  soil,  and 
two  kinds  have  been  described — the  one  kind  converting  ammonia 
into  nitrous  acid,  and  the  other  changing  nitrous  into  nitric  acid. 
To  Winogradsky  and  Frankland  we  are  principally  indebted  for  our 
knowledge  of  these  bacteria. 

Pathogenic  bacteria  are  those  which  are  genetically  related  to 
disease.  Many  organisms  have  been  supposed  to  be  pathogenic,  or 
have  been  described  in  connection  with  diseases,  which  are  only 
saprophytic  associates.  By  saprophytic  we  mean  organisms  which 
feed  upon  dead  organic  matter.  They  include  many  forms  which 
are  found  on  the  skin,  in  the  intestinal  canal,  and  sometimes  in  the 
internal  organs,  especially  the  liver  and  kidneys ;  the  tissues  have 
lost  their  vitality,  and  the  organisms,  through  some  lesion,  have 
been  carried  into  the  circulation. 

That  many  organisms  are  causally  related  to  disease,  there  is 
strong  evidence  in  proof.  No  organism  can  be  considered  to  be  pro- 
ductive of  disease  unless  it  fulfils  the  conditions  which  have  been 
laid  down  by  Koch.  Great  stress  must  be  laid  upon  the  importance 
of  successive  cultivation  through  many  generations,  as  the  objection 
that  a  chemical  virus  may  be  carried  over  from  the  original  source 
is  thus  overcome.  Any  hypothetical  chemical  poison  carried  over 
from  one  tube  to  another  would,  after  a  great  number  of  such 
cultivations,  be  diluted  to  such  an  extent  as  to  be  inappreciable 
arid  absolutely  inert. 

Though  we  may  accept  as  a  fact  the  existence  of  pathogenic 
organisms,  we  are  not  in  all  cases  in  a  position  to  assert  the  means 
by  which  they  produce  their  deleterious  or  fatal  effects.  Many 
theories  have  been  propounded.  It  has  been  suggested  that  the 
pathogenic  organisms  may  be  compared  to  an  invading  army. 
The  cells  or  phagocytes  arrayed  against  them  'endeavour  to  as- 
similate and  destroy  them,  but  perish  themselves  in  the  attempt. 
This  might  explain  the  breaking  down  of  tissue,  and  the  for- 
mation of  local  lesions,  but  does  not  assist  us  in  understanding 
the  fatal  result  in  thirty-six  to  forty-eight  hours  produced  by  the 
inoculation  of  the  bacilli  of  anthrax.  Another  view  is  that  the 
invading  army  seizes  upon  the  commissariat,  appropriating  the 
general  pabulum,  which  is  so  essential  to  the  life  of  the  tissues. 
This  would  hardly  account  for  so  acute  and  fatal  a  result  as  in 
anthrax,  but  would  lead  one  to  expect  symptoms  of  inanition  and 
gradual  exhaustion.  Moreover,  against  this  theory  we  have  the 
fact  that  death  may  result,  in  some  cases,  with  the  presence 


28  BACTERIOLOGY. 

of  comparatively  few  bacilli  in  the  blood ;  and,  again,  the 
blood  may  teem  with  parasites  such  as  the  flagellated  monads  in 
well-nourished,  healthy -looking  rats,  without  apparently  causing 
any  symptoms  whatever.  In  the  same  category  may  be  placed  the 
theory  that  eminently  aerobic  organisms  seize  upon  the  oxygen  of 
the  blood  and  produce  death  by  asphyxia.  Another  explanation  is 
afforded  by  the  suggestion  of  interference  with  the  functions  of  the 
lung  and  kidney  by  mechanical  blocking  of  the  capillaries.  Here 
the  same  objection  is  met  with  in  the  case  of  anthrax,  the  same 
fatal  result  may  occur  with  only  a  few  bacilli,  while  other  cases 
yield  very  beautiful  sections,  looking  like  injected  preparations  from 
the  mapping  out  of  the  capillaries  with  the  countless  crowds  of 
bacilli. 

Putrefactive  bacteria  derive  their  necessary  elements  from  com- 
plex organic  substances,  and  accompanying  the  residue  we  find  the 
presence  of  poisonous  substances.  Pathogenic  bacteria,  in  a  similar 
way,  give  rise  to  virulent  poisons.  Anthrax  bacilli  produce  poisonous 
principles  in  the  blood  which  cause  death,  independently  of  the 
number  of  bacilli,  provided  there  are  sufficient  present  to  develop  a 
fatal  dose. 

It  has  been  also  suggested  that  possibly  a  special  ferment  is 
secreted  by  some  organisms,  and  that  by  the  changes  ultimately 
wrought  by  the  action  of  this  ferment  the  symptoms  and  phe- 
nomena of  disease  arise.  We  have  an  analogy  with  this  theory 
in  the  alkaline  fermentation  of  urine  by  means  of  the  torula  urese.. 
By  the  researches  of  Musculus,  and  later  of  Sheridan  Lea,  it  has 
been  shown  that  a  ferment  is  secreted  by  the  cells  which  can  be 
isolated  in  aqueous  solution,  and  is  capable  of  rapidly  inducing  an 
active  fermentation  of  urea. 

We  can  now  understand  how  it  is  that  in  anthrax  or  in  tuber- 
culosis we  may  find  the  presence  of  only  a  few  bacilli,  or  that  in 
tetanus  we  can  have  such  a  violent  disturbance  of  the  system 
produced  by  the  presence  of  very  few  micro-organisms.  We  may 
conceive  that  different  species  of  bacilli  may  vary  greatly  in  their 
power  of  producing  a  toxin  or  secreting  a  ferment,  just  as  the 
elaboration  of  pigment  is  much  more  marked  in  some  species  than 
in  others ;  thus  it  need  not  follow  that  the  number  of  micro- 
organisms bears  any  relation  to  the  virulence  or  activity  of  the 
.substance  they  produce.  There  is,  however,  yet  another  factor  in 
the  production  of  disease.  We  know  that  in  health  we  are  proof 
against  most  of  these  micro-organisms  ;  if  it  were  not  so,  we  should 
all  rapidly  fall  victims  to  the  tubercle  bacillus  or  others,  w-hich  in 


MORPHOLOGY   AND    PHYSIOLOGY    OF   BACTERIA.  29 

lir.-ilt  li  we  inhale  with  impunity.  We  know  that  a  microbe  may  only 
cause  a  local  lesion  in  one  animal,  but  death  in  another.  It  is  still 
more  striking  that  the  same  micro-organism,  as  is  the  case  with 
anthrax,  may  have  no  effect  whatever  upon  certain  species  of 
animals,  though  it  is  deadly  to  others.  Again,  an  animal  naturally 
susceptible  to  the  effect  of  a  pathogenic  organism  may  be  rendered 
proof  against  it.  These  matters  will  be  discussed  in  a  future 
chapter. 

DISTRIBUTION  OF  BACTERIA. 

Bacteria  are  commonly  described  as  ubiquitous.  They  are  ever 
present  in  the  air,  though  not  in  such  exaggerated  numbers  as  is 
commonly  supposed.  In  nutrient  media  exposed  to  the  air  one  is  often 
astonished  at  times  at  the  comparatively  few  bacteria  which  develop 
in  comparison  to  the  amount  of  floating  matter,  such  as  mineral 
particles,  scales,  spores  of  fungi  and  debris  known  to  be  present. 
In  water  they  are  also  present  in  considerable  numbers,  though  of 
course  varying  according  to  the  character  of  the  water.  Wherever 
there  is  putrefaction,  they  are  present  in  vast  numbers.  In  the  soil, 
in  sewage,  in  the  intestines  and,  in  uncleanly  persons  especially, 
on  the  skin  and  between  the  teeth,  various  species  may  always  be 
found,  but  in  the  healthy  blood  and  healthy  tissues  bacteria  are 
never  present.  In  a  previous  chapter  the  method  of  examining  the 
blood  of  living  persons  has  been  described,  and  there  is,  by  this 
means,  ample  opportunity  for  satisfying  oneself  that  bacteria  are 
never  to  be  found  in  the  blood  in  health.  The  organs  remove^  from  a 
perfectly  healthy  animal,  with  the  necessary  precautions,  and  placed 
in  sterilised  media,  can  be  kept  indefinitely  without  undergoing 
putrefaction,  or  giving  any  development  of  bacteria.  This  has  been 
established  by  many  observers,  notably  Cheyne  and  Hauser ;  and 
the  results  of  former  observers  to  the  contrary  must  be  attributed 
to  imperfect  methods  admitting  of  accidental  contamination. 


CHAPTER    III. 

EFFECT   OF   ANTISEPTICS   AND    DISINFECTANTS   ON    BACTERIA. 

IN  the  previous  chapter  several  conditions  were  alluded  to  which 
affected  the  growth  of  bacteria,  such  as  the  nature  of  the 
nutrient  soil,  temperature,  light,  and  electricity.  The  effect  of 
certain  chemical  substances,  and  of  excessive  heat  and  cold,  was 
also  mentioned ;  but  this  constitutes  a  subject  of  such  practical 
importance  that  it  must  be  considered  more  fully. 

Agents  which  retard  the  growth  of  bacteria  are  generally  spoken 
of  as  antiseptics,  as  distinguished  from  disinfectants  which  altogether 
destroy  their  vitality. 

Though  chemical  disinfectants,  or  germicides,  when  diluted,  act 
as  efficient  antiseptics,  the  converse,  that  an  antiseptic  in  a  suffi- 
ciently concentrated  form  will  act  as  a  disinfectant,  is  not  the  case. 
The  term  "  antiseptic,"  indeed,  should  be  restricted  to  those  sub- 
stances or  agents  which  arrest  the  changes  bacteria  produce,  but 
which  do  not  prevent  their  springing  into  activity  when  removed 
to  favourable  conditions.  Thus  excessive  heat,  which  destroys 
bacteria  and  their  spores,  is  a  true  disinfectant ;  and  excessive  cold, 
which  only  benumbs  them,  retarding  their  development  without 
killing  them,  is  an  antiseptic. 

Spores  have  a  greater  power  of  resisting  the  action  of  these 
various  agents  than  the  parent  cells,  and  many  species  of  micro- 
organisms differ  from  each  other  in  their  resisting  power.  An 
exact  knowledge  of  the  subject  can,  therefore,  only  be  based  upon 
investigations  which  will  determine  the  effect  of  these  agents  upon 
pure  cultivations  of  the  different  micro-organisms  causally  related  to 
putrefaction  and  disease.  In  the  latter  case,  especially,  this  is  not 
possible  in  the  present  state  of  our  knowledge.  In  some  cases  of 
communicable  disease  there  is  considerable  doubt  as  to  the  etiological 
importance  of  the  organisms  which  have  been  described;  in  other 
cases  no  organisms  have  as  yet  been  discovered,  or  the  organisms 

30 


EFFECT    OF   ANTISEPTICS   AND    DISINFECTANTS   ON    BACTERIA.       31 

cannot  be  artificially  cultivated,  or  the  disease  is  not  reproduced  by 
inoculation,  so  that  there  is  no  means  of  testing  whether  the  agents 
have  had  any  effect.  One  can,  therefore,  only  draw  general 
conclusions  by  selecting  some  well-known  pathogenic  and  non- 
pathogenic  micro-organisms,  and  considering  the  influence  of 
chemicals,  of  hot  air  and  of  steam  upon  them,  as  representing  the 
•effect  upon  the  various  contagia  of  disease  and  the  causes  of 
putrefaction. 

Such  knowledge  must  necessarily  prove  of  the  greatest  im- 
portance :  to  the  sanitarian,  who  is  concerned  in  preventing  the 
spreading  of  disease  and  in  the  disposal  of  putrefactive  matter;  to 
the  surgeon,  who  is  anxious  to  exclude  micro-organisms  during 
.surgical  operations,  and  to  arrest  the  development  of  bacteria  which 
have  already  gained  an  entrance  in  wounds ;  to  the  physician,  in 
the  treatment  of  micro-parasitic  diseases. 

The  sanitarian  and  the  surgeon  must  profit  directly  by  such 
•experiments,  for  in  the  disinfection  of  clothes  and  the  sick-room  by 
the  one,  and  in  the  application  of  antiseptic  dressings  and  lotions 
by  the  other,  the  micro-organisms  are  encountered,  as  in  the  experi- 
ments, outside  the  living  body. 

The  physician,  on  the  other  hand,  is  principally  concerned  in 
dealing  with  micro- parasites  when  circulating  in  the  blood,  or 
•carrying  on  their  destructive  processes  in  the  internal  tissues.  So 
far  as  our  knowledge  at  present  goes,  the  physician  can  avail  him- 
.self  but  little  of  the  effect  of  the  direct  application  of  the  substances 
which  have  been  found  to  retard  or  destroy  the  growth  of  the 
organisms  in  artificial  cultivations,  for  the  concentrated  form  in 
which  they  would  have  to  be  administered  would  prove  as  /  deleteri- 
ous or  as  fatal  to  the  host  as  to  the  parasites.  Thus  Koch  has 
stated  that  to  check  the  growth  of  the  anthrax  bacillus  in  man  it 
would  be  necessary  that  there  should  be  twelve  grammes  of  iodine 
•constantly  in  circulation,  and  that  the  dose  of  quinine  necessary 
to  destroy  the  spirilla  of  relapsing  fever  would  be  from  twelve  to 
sixteen  grammes.  The  retarding  influence,  however,  of  certain 
substances  when  diluted,  and  the  fact  that  disinfectants  are  some- 
times equally  efficacious  in  a  diluted  form  when  their  application  is 
prolonged,  seem  to  indicate  measures  which  may  be  adopted,  in  some 
«ases,  with  chances  of  success,  such  as  the  inhalation  of  antiseptic 
vapours  in  phthisis.  For  the  most  part  the  physician  must  look 
rather  to  combating  the  effects  of  micro-organisms  by  restoring  to 
its  normal  standard  the  lowered  vitality  which  enabled  the  bacteria 
to  get  a  footing. 


32  BACTERIOLOGY. 

There  is  no  wider  field  for  research  than  the  determination  of 
the  real  effect  of  disinfectants  and  antiseptics.  Painstaking  and 
laborious  as  the  researches  are  which  have  been  hitherto  made,  the- 
subject  is  so  beset  with  fallacies,  leading,  in  some  cases,  to  totally 
erroneous  conclusions,  that  it  is  not  surprising  that  one  meets  on 
all  sides  with  conflicting  statements.  The  author  has  no  intention 
of  analysing  these  results,  but  a  general  idea  will  be  given  of  the- 
methods  which  have  been  employed,  and  for  further  details  reference 
must  be  made  to  the  original  papers  mentioned  in  the  bibliography. 

Chemical  Substances. — It  was  customary  to  judge  of  the  power 
of  a  disinfectant  or  antiseptic  by  adding  it  to  some  putrescent  liquid, 
A  small  portion  of  the  latter  was,  after  a  time,  transferred  to 
some  suitable  nourishing  medium,  and  the  efficacy  of  the  substance 
estimated  by  the  absence  of  cloudiness,  odour,  or  other  sign  of 
development  of  bacteria  in  the  inoculated  fluid.  Koch  pointed  out 
the  errors  that  might  arise  in  these  experiments  from  accidental 
contamination,  or  from  there  being  no  evidence  of  the  destruction 
of  spores,  and  we  are  indebted  to  him  for  a  complete  and  careful 
series  of  observations  with  more  exact  methods. 

Instead  of  employing  a  mixture  of  bacteria,  Koch's  plan  was  to 
subject  a  pure  cultivation  of  some  well-known  species  with  marked 
characteristics  to  the  reagent  to  be  tested.  A  small  quantity  was 
then  transferred  to  fresh  nourishing  soil,  under  favourable  con- 
ditions, side  by  side  with  nutrient  material  inoculated  from  a 
cultivation  without  treatment  with  the  disinfectant.  The  latter 
constituted  a  control  test,  which  is  most  essential  in  all  such 
experiments.  To  test  the  resistant  power  of  bacteria  which  are 
easily  destroyed,  two  species  were  selected,  Micrococcus  prodigiosusr 
and  the  bacillus  of  blue  pus.  These  were  cultivated  on  potatoes, 
the  surfaces  of  which  were  sliced  off  and  dried.  A  fragment  trans- 
ferred to  freshly  prepared  potato  gave  rise  to  a  growth  of  the 
particular  micro-organism  ;  but  if  after  treatment  with  some  reagent 
no  growth  occurred,  the  conclusion  was  drawn  that  the  reagent  was 
efficacious  in  destroying  the  vitality  of  the  bacteria. 

Anthrax  bacilli  in  blood,  withdrawn  from  an  animal  just  killed, 
were  taken  to  represent  sporeless  bacteria,  while  silk  threads  steeped 
in  an  artificial  cultivation  of  the  bacilli  and  dried,  afforded  a  means 
of  testing  the  vitality  of  spores. 

Even  by  employing  pure  cultivations  on  solid  media,  great 
precautions  were  necessary  to  avoid  mistakes.  When,  for  instance, 
a  large  quantity  of  the  growth  which  had  been  subjected  to  some 
chemical  solution  was  carried  over  to  the  fresh  tube  containing 


EFFECT   OF   ANTISEPTICS   AND    DISINFECTANTS   ON    BACTERIA.       33 

the  nutrient  medium,  or  when  a  silk  thread,  which  had  been  dipped 
in  a  solution,  was  directly  transferred  to  the  new  soil,  enough  of  the 
supposed  disinfectant  might  be  mechanically  carried  over  to  retard 
the  development  of  the  bacteria,  though  it  was  ineffectual  in 
destroying  them.  From  a  growth  not  appearing,  it  was  concluded 
that  the  spores  or  the  bacteria  had  been  affected,  and  so  a 
mistake  occurred.  To  avoid  this,  Koch  made  a  point  of  transfer- 
ring a  minimum  of  the  disinfected  growth  to  as  large  a  cultivation 
area  as  possible,  so  that  any  chemical  substance  mechanically 
carried  over  would  be  so  diluted  as  to  be  inert.  For  the  same 
reason,  threads,  after  withdrawal  from  the  disinfecting  solution, 
were  rinsed  in  sterilised  water,  or  weak  alcohol,  and  then  trans- 
planted ;  or,  instead  of  judging  from  the  development  on  nutrient 
gelatine,  the  effect  of  inoculation  in  a  healthy  animal  was  made 
the  test. 

A  few  examples  may  be  quoted  in  illustration.  Silk  threads, 
impregnated  with  anthrax  spores,  were  placed  in  bottles  containing 
carbolic  acid  of  various  strengths.  A  thread  was  removed  from  each 
on  successive  days,  and  transferred  to  nutrient  gelatine,  and  the 
result  noted.  It  was  found  that  immersion  of  the  thread  in  a  5  per 
cent,  solution  of  carbolic  acid  was  sufficient  in  two  days  to  effect 
complete  sterilisation,  and  seven  days  in  a  3  per  cent,  solution  was 
equally  efficacious.  Since  for  practical  purposes  a  strength  should 
be  selected  which  would  be  effectual  in  twenty-four  hours,  Koch 
recommended  that  for  general  use,  allowing  for  deterioration  by 
keeping,  a  solution  containing  not  less  than  5  per  cent,  should  be 
employed,  and  for  complex  fluids  probably  a  still  higher  percentage 
would  be  necessary.  In  the  case  of  sporeless  bacilli  the  results  were 
very  different.  Blood  containing  the  bacilli,  from  an  animal  just 
killed,  was  dried  on  threads,  and  after  exposure  for  two  minutes  to 
a  1  per  cent,  solution,  was  completely  sterilised;  and  fresh  blood 
mixed  with  a  1  per  cent,  carbolic  solution  produced  no  effect  when 
inoculated.  On  the  other  hand,  when  the  blood  was  mixed  with  a 
•5  per  cent,  solution,  the  virulence  was  not  destroyed.  The  facility 
with  which  the  bacilli  are  destroyed,  compared  with  their  spores, 
illustrates  how  easily  errors  may  occur,  when  mere  arrest  of  growth 
or  loss  of  motility  is  regarde_d  as  a  sign  of  the  efficacy  of  disinfection. 

To  test  vapours,  Koch  exposed  anthrax  spores  or  the  spores 
which  occur  in  garden  earth  by  suspending  them  over  solutions, 
such  as  bromine  or  chlorine,  in  a  closed  vessel.  After  a  time  they 
were  transferred  to  a  nutrient  medium  to  test  their  vitality.  To 
test  the  power  of  sulphurous  acid  gas,  the  spores  were  spread  about 

3 


34  BACTERIOLOGY. 

in  a  room  in  which  the  gas  was  generated  by  burning  sulphur  in 
the  ordinary  way  for  disinfecting  a  room.  To  test  chemicals  which 
might  be  recommended  for  disinfecting  vans  and  railway  carriages, 
spores. were  laid  on  boards,  which  were  then  washed  or  sprayed,  and 
the  spores  then  transferred  to  the  nutrient  gelatine. 

Sternberg  has  also  made  an  elaborate  series  of  experiments  with 
regard  to  the  action  of  germicides.  In  this  case  cultivations  of 
well-known  pathogenic  organisms  in  liquid  media  were  employed. 
The  supposed  germicide  was  added  to  the  liquid  cultivation,  and 
after  two  hours  a  fresh  flask  of  sterilised  culture  was  inoculated  from 
the  disinfected  cultivation,  and  placed  in  the  incubator.  In  twenty- 
four  to  forty-eight  hours,  if  the  chemical  proved  inefficient,  there 
was  evidence  of  a  growth  of  bacteria.  Blyth  has  investigated  the 
disinfection  of  cultivations  of  Bacterium  termo,  of  sewage,  and 
typhoid  excreta,  and,  in  conjunction  with  Klein,  the  effect  of  well- 
known  disinfectant  materials  on  anthrax  spores.  Miquel,  Laws, 
and  others  have  also  contributed  to  our  knowledge  of  the  effect 
of  antiseptics  and  disinfectants  upon  micro-organisms.  In  spite  of 
all  that  has  be'en  done  there  is  room  for  many  workers ;  a  great 
deal  of  ground  must  be  gone  over  again  to  rectify  discrepancies, 
examine  conflicting  results,  and  thus  determine  what  observations 
may  be  relied  upon  for  practical  application. 

This  may  be  illustrated  by  referring  in  detail  to  some  experiments 
made  with  corrosive  sublimate.  Koch  investigated  a  long  list  of 
chemical  reagents,  and  according  to  these  experiments  the  salts  of 
mercury,  and  the  chloride  especially,  proved  most  valuable.  Where 
heat  is  not  admissible,  these  compounds  were  therefore  highly 
recommended,  though  their  poisonous  nature  is  a  drawback  to  their 
indiscriminate  use.  Koch  stated  that  for  disinfecting  a  ship's  bilge, 
where  a  5  per  cent,  solution  of  carbolic  acid  must  be  left  forty-eight 
hours,  a  1  in  1000  solution  of  mercuric  chloride  would  only  require 
a  few  minutes. 

But  there  was  good  reason  for  doubting  the  efficacy  of  very  dilute 
solutions;  for,  though  according  to  Koch's  experiments  anthrax 
spores  subjected  to  a  1  in  20.000  solution  of  mercuric  chloride  for 
ten  minutes,  and  then  washed  in  alcohol,  gave  no  growth  in  nutrient 
gelatine,  silk  threads  exposed  for  ten  minutes  to  a  1  in  20,000 
solution,  or  even  1  in  10,000,  still  proved  fatal  to  mice. 

Herroim  cultivated  ordinary  septic  bacteria  in  albuminous 
nitrates,  containing  1  in  2000,  and  concluded  that  the  value  of 
mercuric  chloride  as  an  antiseptic  was  much  over-rated.  It  is  pre- 
cipitated by  albumins  though,  as  Lister  has  shown,  the  precipitate 


EFFECT   OF   ANTISEPTICS   AND    DISINFECTANTS   ON    BACTERIA.       35 

of  albuniinate  of  mercury  is  redissolved  when  there  is  an  excess  of 
albumin  present. 

Geppert,  and  later  Behring,  recognised  that  the  methods  employed 
for  testing  the  efficacy  of  corrosive  sublimate  were  unreliable.  They 
found,  for  example,  that  corrosive  sublimate  could  not  be  removed 
from  silk  threads  by  washing ;  and  therefore  to  study  the  effect  of 
this  antiseptic  acting  for  a  given  time,  it  was  necessary  to  dip  the 
threads  in  ammonium  sulphide  solution  after  the  treatment  with 
corrosive  sublimate. 

The  author  confirmed  the  results  of  Geppert  and  Behring,  and 
made  a  series  of  experiments  to  test  the  value  respectively  of  carbolic 
acid  and  corrosive  sublimate  in  antiseptic  surgery.  The  method 
of  dipping  an  infected  thread  into  an  antiseptic  solution  for  a  few 
minutes,  and  then  transferring  it  to  the  surface  of  a  nutrient  medium 
to  test  its  efficacy  in  a  given  time,  was  discarded  as  fallacious ;  the 
thread  being  still  wet  with  the  solution  when  transferred  to  the 
medium,  it  was  obvious  that  the  action  of  the  antiseptic  continued 
for  many  days.  To  wash  infected  silk  threads  with  alcohol  after 
exposure  to  the  antiseptic  to  stop  its  further  action,  also  proved  to 
be  a  fallacious  method,  for  the  author  found  in  control  experiments 
that  absolute  alcohol  will  destroy  Streptococcus  pyogenes,  erysipelatis, 
arid  Staphylococcus  pyogenes  aureus,  acting  for  only  one  minute. 
Other  methods  were  therefore  resorted  to,  and  cultures  on  the 
sloping  surface  of  nutrient  agar  were  at  first  used.  The  antiseptic- 
was  poured  into  the  culture  tube  until  the  growth  was  covered, 
and  when  it  had  acted  for  a  definite  time  (one  minute,  five  minutes, 
or  fifteen  minutes)  a  solution  was  added  which  immediately  stopped 
further  action.  In  the  case  of  corrosive  sublimate,  ammonium  sul- 
phide was  employed,  which  is  quite  inert  as  an  antiseptic.  The  liquid 
contents  of  the  test  tube  were  carefully  poured  off,  and  an  inoculation 
was  made  into  a  fresh  tube  of  broth  or  agar  from  the  culture  still 
adhering  to  the  surface  of  the  nutrient  medium.  As  the  results 
disproved  the  efficacy  of  corrosive  sublimate,  it  was  thought  possible 
that  the  solution  had  not  been  able  in  the  time  to  penetrate  the 
film  of  growth.  Another  plan  was  accordingly  adopted.  Cultures 
were  made  in  broth,  and  when  fully  developed  the  supernatant  liquid 
was  carefully  poured  off.  Corrosive  sublimate  solution  was  added  to 
the  test  tube,  and  agitated  until  any  flocculent  masses  were  dis- 
integrated and  the  whole  of  the  liquid  became  uniformly  turbid. 
Ammonium  sulphide  was  added  when  the  time  had  expired,  and 
tubes  of  fresh  broth  were  inoculated  with  the  mixture.  In  the  case 
of  carbolic  acid  the  cultures,  after  its  action,  were  thoroughly  washed 


36  BACTERIOLOGY. 

with  water,  and  its  efficacy  tested  by  making  inoculations  from  the 
cultures  in  fresh  media.  The  results  were  entirely  in  favour  of 
carbolic  acid.  Staphylococcus  pyogenes  aureus  and  Streptococcus 
pyogenes  were  not  destroyed,  even  when  corrosive  sublimate  solution 
of  1  in  1000  was  allowed  to  act  for  an  hour.  In  the  case  of  the 
cultures  of  streptococcus  of  erysipelas  the  results  were  different. 
A  solution  of  1  in  10,000  had  no  effect,  but  1  in  4,000,  acting  for 
one  minute,  destroyed  the  culture.  With  carbolic  acid  the  results 
were  very  striking.  Cultures  were  exposed  to  solutions  of  1  in  20, 
1  in  30,  1  in  40,  1  in  50,  for  one  minute,  five  minutes,  fifteen 
minutes.  The  attempts  to  make  subcultures  in  every  case  failed. 
Carbolic  acid  I  in  40,  acting  for  only  one  minute,  was  sufficient  to 
destroy  Streptococcus  pyogenes  and  Streptococcus  erysipelatis  and 
Staphylococcus  pyogenes  aureus.  .Further  experiments  were  made 
with  tubercular  sputum,  the  test  being  subsequent  inoculation  of 
guinea-pigs.  Corrosive  sublimate  solution  as  strong  as  1  in  500  had 
no  effect,  but  1  in  20  carbolic  acid,  shaken  up  with  the  sputum  for 
one  minute,  completely  neutralised  it. 

Koch's  statements  with  reference  to  the  germicidal  power  of 
corrosive  sublimate  in  extremely  weak  solutions  had  led  Lister  to 
substitute  it  for  carbolic  acid  as  a  detergent  in  surgery.  The  author's 
experiments,  which  were  undertaken  in  1892,  encouraged  Lister  to 
revert  to  the  use  of  carbolic  acid,  which,  indeed,  had  always  proved 
efficacious  in  surgical  practice.  Lister  pointed  out  that  carbolic  acid 
has  also  the  great  advantage  of  combining  eagerly  with  fats  and 
epidermis,  so  that  the  seat  of  operation  can  be  effectually  cleansed. 

These  experiments  also  point  to  the  conclusion  that  carbolic  acid 
should  be  used  in  hospital  wards  for  the  disinfection  of  tubercular 
sputum  instead  of  mercuric  chloride  and  other  less  efficacious  dis- 
infectants commonly  in  use. 

Hot  Air  and  Steam. — Koch,  in  conjunction  with  Wolfhiigel,  also 
made  exhaustive  experiments  to  test  the  value  of  hot  air.  A  similar 
plan  was  adopted  to  that  employed  in  disinfection  with  chemicals. 
Bacteria  and  spores  were  subjected  for  a  certain  time  to  a  known 
temperature  in  the  hot-air  chamber,  and  then  were  transferred  to  a 
nourishing  soil  or  inoculated  in  animals. 

Paper  parcels,  blankets,  bags,  and  pillows,  containing  samples  of 
micro-organisms  wrapped  up  inside,  were  also  placed  in  the  hot-air 
chamber,  to  test  the  power  of  penetration  of  heat. 

The  conclusions  from  these  experiments  were  as  follows  : — 

Sporeless  micro-organisms  at  a  little  over  100°  C.  are  destroyed 
in  one  hour  and  a  half. 


EFFECT   OF   ANTISEPTICS   AND    DISINFECTANTS   ON    BACTERIA.      37 

Spores  of  bacilli  require  three  hours  at  140°  C. 

If  enclosed  in  pillows  and  blankets,  exposure  from  three  to  four 
hours  to  140°  C.  is  necessary. 

Spores  of  fungi  require  one  and  a  half  hours  at  110°  C.  to 
115°  C. 

Further  experiments  showed  that  at  the  temperature  necessary 
for  the  destruction  of  spores  of  bacilli  almost  all  fabrics  are  more  or 
less  injured. 

Koch,  in  conjunction  with  Gaffky  and  Loffler,  also  investigated 
the  effect  of  steam  under  pressure  and  at  the  atmospheric  pressure. 

Rolls  of  flannel  with  anthrax  spores  or  earth  spores,  and  a 
thermometer  wrapped  up  inside,  were  subjected  to  steam,  and 
the  results  compared  with  the  effect  obtained  with  hot  air. 

Thus  in  hot  air  four  hours'  exposure  to  a  temperature  of  130°  C. 
to  140°  C.  brought  the  temperature  inside  the  roll  to  85°  C.,  and  the 
spores  were  not  injured ;  on  the  other  hand,  exposure  to  steam 
under  pressure  at  120°  C.  for  one  and  a  half  hours,  raised  the 
internal  temperature  to  117°  C.  and  killed  the  spores. 

By  such  experiments  the  superior  penetrative  power  of  steam- 
heat  was  established. 

To  test  stsam-heat  at  the  atmospheric  pressure,  water  was  boiled 
in  a  glass  flask  with  its  neck  prolonged  by  means  of  a  glass  tube,  the 
temperature  ia  which  was  found  to  be  uniform  throughout.  Anthrax 
and  earth  spores  placed  in  the  tube  were  found  to  be  unable  to  with- 
stand steam  at  100°  C.  even  for  a  few  minutes.  It  was,  therefore, 
concluded  that  disinfection  by  steam  at  atmospheric  pressure  was 
superior  to  hot  air  from  its  greater  efficiency,  and  to  steam  under 
pressure  from  the  simplicity  of  the  necessary  apparatus. 

Parsons  and  Klein  made  some  experiments  which  were  more 
in  favour  of  dry  heat  than  the  above.  These  observers  state  that 
anthrax  bacilli  are  destroyed  by  an  exposure  of  five  minutes  at 
from  100°  C.  to  103°  C.  and  that  anthrax  spores  are  destroyed 
in  four  hours  at  104°  0.,  or  in  one  hour  at  118°  C.  Guinea-pigs 
inoculated  with  tuberculous  pus  which  had  been  exposed  for  five 
minutes  to  104°  C.,  remained  unaffected.  They  concluded  that  as 
none  of  the  infectious  diseases,  for  which  disinfecting  measures 
are  in  practice  commonly  applied,  are  known  to  depend  upon  the 
presence  of  bacilli  in  a  spore -b3aring  condition,  their  contagia 
are  not  likely  to  retain  their  activity  after  being  heated  for  an 
hour  to  105°  C.  (220°  Fahr.) 

In  experiments  with  steam  the  results  were  in  accordance 
with  those  already  given,  and  complete  penetration  of  an  object 


38  BACTERIOLOGY. 

by  steam-heat  for  more  than  five  minutes  was  deemed  sufficient. 
They  also  arrived  at  the  same  result  as  in  Koch's  experiments, 
viz.,  that  steam-chambers  are  preferable  to  those  in  which  dry  heat 
is  employed,  though  it  must  be  borne  in  mind  that  some  articles, 
such  as  leather,  are  injured  by  exposure  to  steam. 

PRACTICAL  APPLICATION. 

Nurses  and  others  attending  infectious  cases  should  freely  use 
1  in  40  carbolic  for  the  hands  and  a  weaker  solution  for  the  body 
generally.  The  skin  of  patients  after  recovery  should  be  sponged 
with  1  in  40  carbolic.  The  dead  should  be  wrapped  up  in  a  sheet 
soaked  in  1  in  20  carbolic  acid  or  a  strong  solution  of  chloride  of 
lime.  Infected  clothing  and  bedding  should  be  burnt  unless  in  excep- 
tional cases,  when  they  may  be  disinfected  by  boiling,  or  by  exposure 
to  dry  heat  at  105°C.  to  110°C.  for  three  hours,  or  by  steaming 
at  100°C.  for  fifteen  minutes.  Leather  and  other  articles  which 
would  be  destroyed  by  any  of  these  processes  should  be  sponged  with 
1  in  40  carbolic.  The  walls  of  the  sick-room  and  furniture  should 
be  exposed  to  the  fumes  of  burning  sulphur,  and  next  day  washed 
down  with  1  in  40  carbolic,  and  the  room  freely  ventilated  by 
opening  all  windows  and  doors.  Rags  should  be  burnt,  or  dis- 
infected by  boiling  or  exposure  to  steam  when  supplied  to  manu- 
facturers. The  importation  of  rags  from  places  where  there  are 
cases  of  cholera  or  small-pox  should  be  prohibited.  Infected  ships 
must  be  fumigated  with  sulphur,  and  the  bilge  disinfected  with 
carbolic  acid.  Infected  railway  carriages  should  be  disinfected  in 
the  same  way  as  a  sick-room. 

Tubercular  sputum,  cholera  and  typhoid  evacuations  and  other 
excreta  should  be  disinfected  by  1  in  20  carbolic  acid,  or  by  a  strong 
solution  of  chloride  of  lime. 


CHAPTER    IV. 

CHEMICAL    PRODUCTS   OF    BACTERIA. 

THE  products  of  the  metabolism  induced  by  bacteria  may  be  divided 
into  three  classes  :  (1)  ptomaines  or  alkaloids  ;  (2)  albumoses  or  tox- 
albumins  ;  and  (3)  enzymes.  Alkaloids  and  albumoses  are  directly 
poisonous ;  enzymes  or  ferments  are  harmless  except  in  the  presence 
of  proteids,  which  they  are  capable  of  transforming  into  poisonous 
albumoses. 

PTOMAINES  AND  Tox- ALBUMINS. 

The  study  of  these  products  may  be  said  to  date  back  to  1822, 
when  Gaspard  and  Stick  found  an  intensely  poisonous  principle 
in  cadaverous  extracts.  In  1856  Panum  discovered  a  poisonous 
substance  in  putrid  flesh;  and  in  1863  Bergmann  and  Smiedeberg 
found  a  nitrogenous  crystallisable  substance  in  putrid  beer  which 
they  named  sepsin.  In  1872  Gautier  found  that  the  decomposition 
of  fibrine  led  to  the  formation  of  various  complex  alkaloidal  sub- 
stances, and  in  1875  Richardson  obtained  in  pyaemia  an  alkaloid, 
septin.  This  subject,  however,  received  most  attention  from  the 
classical  researches  of  Selmi,  the  Italian  toxicologist.  Selmi,  in  a 
celebrated  poisoning  case,  demonstrated  the  presence  of  an  alkaloid 
as  the  result  of  post-mortem  changes.  Similar  substances  were 
found  in  alcohol  in  which  morbid  specimens  had  been  preserved. 
Thus  the  researches  of  Gautier  and  Selmi  established  the  fact  that 
albuminoid  material  undergoing  decomposition  leads  to  the  forma- 
tion of  cadaveric  alkaloids.  These  animal  alkaloids  Selmi  named 
ptomaines.  Brieger,  finding  the  bases  derived  from  the  products  of 
putrefaction  less  poisonous  than  those  obtained  from  the  pathogenic 
bacteria,  suggested  the  term  toxins  for  the  latter.  Ptomaines  have 
been  divided  into  two  classes — those  which  are  non- oxygenous,  liquid, 
and  volatile,  and  those  which  are  oxygenous,  solid,  and  crystallisable. 
They  are,  for  the  most  part,  precipitated  by  the  ordinary  reagents 

39 


40  BACTERIOLOGY. 

for  alkaloids,  such  as  chloride  of  gold,  double  iodide  of  mercury 
and  potassium,  picric  acid,  and  tannin.  Phospho-molybdic  acid 
precipitates  them  without  exception.  They  are  powerful  reducing 
agents.  Ferro- cyanide  of  potassium  is  converted  into  ferri-cyanide  in 
their  presence,  and  the  addition  of  ferric  chloride  gives  the  Prussian 
blue  test.  Selini  discovered  this  test,  and  Brouardel  and  Boutmy 
regarded  it  as  absolutely  characteristic  of  ptomaines ;  but  this  is 
not  the  case  ;  some  vegetable  alkaloids,  for  example,  behave  in  the 
same  way. 

As  examples  of  the  non-oxygenous  ptomaines  there  are  : — 

Parvolin  (C9H13N)  an  oily  base  of  an  amber  colour  prepared 
from  putrid  mackerel  and  horse-flesh. 

Hydrocollidin  (C8H13N),  from  the  same  source.  It  is  highly  toxicT 
being  compared  by  Gautier  to  the  venom  of  the  cobra  di  capello. 

Collidin  (C8!!11^),  from  putrid  gelatine  and  the  pancreas  of  a 
bullock,  also  highly  toxic. 

Neuridin  (C5H14N2),  from  fish,  flesh,  and  decaying  cheese. 

Saprin  (C5H14N2),  isomeric  with  neuridin. 

Cadaverin  (C5H14N2),  a  third  isomeride,  from  ordinary  putrefac- 
tion and  herring  brine. 

Putrescin  (C4H12N2)  from  putrefaction. 

The  oxygenous  ptomaines  are  in  some  instances  found  also  in 
healthy  tissues.  They  include  the  following  : — 

Neurin  (C5H13NO),  found  in  cadaveric  putrefaction. 

Cholin  (C5H5N02),  in  bile. 

Muscarin  (C5H13N02),  in  a  poisonous  mushroom,  Agaricus  mus- 
carius,  and  in  putrid  fish.  These  are  all  highly  poisonous. 

Gadinin  (C7H16N02),  in  putrefying  codfish. 

Mytilotoxm  (C6H15N02),  in  poisonous  mussels. 

Poisonous  alkaloids  are  of  great  importance  in  connection  with 
those  cases  of  meat  poisoning  produced  by  sausages,  hams,  poultry, 
and  cheese.  Tyrotoxicon  is  a  poisonous  alkaloid  obtained  from  cheese. 

The  toxic  substances  of  most  interest  to  the  bacteriologist  are 
those  isolated  from  pure  cultivations  of  pathogenic  bacteria,  such  as 
typhotoxin,  isolated  by  Brieger  from  cultivations  of  the  bacillus  of 
typhoid  fever,  and  tetanin,  from  cultivations  of  the  tetanus  bacillus  ; 
and  the  poisons  known  as  albumoses  or  tox-albumiris,  which  are 
allied  to  the  albuinose  of  snake  poison. 

Pasteur,  in  1885,  suggested  that  in  anti-rabic  inoculations  the 
immunity  resulted  from  the  action  of  a  substance  secreted  by  a 
microbe,  though  the  microbe  has  not  as  yet  been  discovered  in 
rabies.  Salmon  produced  immunity  from  hog  cholera  by  the  injec- 


CHEMICAL   PRODUCTS   OF   BACTERIA.  41 

tion  of  the  toxic  products  in  filtered  culture  fluids.  Wooldridge, 
Hankin,  and  Martin  studied  the  products  of  Bacillus  anthracis. 
Charrin,  and  later  Woodhead,  Wood,  and  Blagovestchensky,  investi- 
gated on  these  lines  Bacillus  pyocyaneus.  Roux  and  Chamberland 
experimented  with  the  bacillus  of  malignant  redema;  Roux  with 
symptomatic  anthrax;  Chantemesse  and  Widal  with  the  typhoid 
bacillus.  Roux,  Yersin,  Brieger,  Fraiikel,  Martin,  and  Behring 
worked  on  the  same  lines  with  diphtheria.  Koch  introduced 
tuberculin,  Kalning  mallein,  while  others  have  utilised  the  products 
of  streptococci  and  pneumococci.  Anrep  found  an  albumose  in  the 
medulla  of  rabid  animals,  and  Babes  claims  to  have  found  an 
albumose  in  both  rabies  and  glanders. 

Cholera. — Brieger  found  several  ptomaines,  including  putrescin 
and  cadaverin,  in  pure  cultures  of  the  spirillum  of  Asiatic  cholera, 
and  Petri  found  in  addition  to  poisonous  bases  a  proteid  body  which 
produces  in  guinea-pigs  muscular  tremors,  paralysis,  and  a  rapidly 
fatal  result.  Roux  and  Yersin  obtained  from  cultures  a  tox-albumin 
insoluble  in  water,  which  kills  guinea-pigs  in  two  or  three  days, 
but  has  no  effect  on  rabbits.  Pfeiffer  also  investigated  the  toxic 
substances  in  cultures.  Chloroform,  thymol,  and  drying  destroyed 
comma -bacilli,  leaving  their  toxic  products  unaffected.  Concentrated 
solutions  of  neutral  salts  and  boiling  produced  secondary  toxic 
substances,  but  the  original  toxic  substances  were  ten  or  twenty 
times  more  virulent. 

Typhoid  Fever.— Typhotoxiu  (C7H17N02),  the  alkaloid  ob- 
tained by  Brieger  from  cultures  of  the  typhoid  fever  bacillus,  produces 
in  mice  and  guinea-pigs  salivation,  rapid  breathing,  dilatation  of  the 
pupil,  diarrhoea,  and  death  in  twenty-four  to  forty-eight  hours.  At 
the  post-mortem  examination  the  heart  is  found  in  a  state  of  systolic 
contraction,  and  the  condition  of  the  heart  after  death  and  the 
absence  of  convulsions  during  life  serve  to  distinguish  typhotoxin 
from  an  isomeric  base  obtained  by  Brieger  from  putrid  horse-flesh. 
Roux  and  Yersin  have  obtained  a  tox-albumin.  It  is  soluble  with 
difficulty  in  water,  arid  more  toxic  to  rabbits  than  guinea-pigs. 

Tetanus. — Brieger  obtained  the  alkaloid  tetanin  from  impure 
cultures  of  the  tetanus  bacillus.  It  is  a  base  having  the  formula 
C13H'22N204.  The  hydrochloride  is  a  very  deliquescent  salt,  and 
soluble  in  alcohol.  Tetanin  injected  into  guinea-pigs  produces 
rapid  breathing,  followed  by  tetanic  convulsions.  Another  toxic 
product,  tetanotoxin  (C5IinN),  produces  the  same  effects  as  tetanin. 
The  formula  of  a  third  base,  spasmotoxin,  has  not  been  determined. 
Cadaverin  and  putrescin  are  also  present  in  cultures.  Kitasato  and 


42  BACTERIOLOGY. 

Weyl  analysed  the  products  of  pure- cultures,  and  obtained  the  same 
substances,  tetanin  and  tetanotoxin ;  and  subsequently  Brieger  and 
Frankel  found  that  in  pure-cultures  a  tox- albumin  could  be 
obtained  which  is  soluble  in  water,  and  infinitely  more  active  than 
the  toxic  ptomaines. 

Anthrax. — In  1887  Wooldridge  succeeded  in  protecting  rabbits 
from  anthrax  by  a  new  method.  A  proteid  body  obtained  from  the 
testis  and  from  the  thyrnus  gland  was  used  as  the  culture  fluid. 
This  proteid  substance  was  dissolved  in  dilute  alkali,  and  the  solution 
sterilised  by  repeated  boiling.  This  was  inoculated  with  the  anthrax 
bacillus,  and  kept  at  37°  C.  for  two  or  three  days.  A  small  quantity 
of  the  filtered  culture  fluid  injected  into  the  circulation  in  rabbits 
produced  immunity  from  anthrax.  Subcutaneous  inoculation  of 
extremely  virulent  anthrax  blood,  made  simultaneously  with  the  injec- 
tion of  the  protecting  fluid,  produced  no  effect.  Wooldridge  showed 
that  the  growth  of  the  anthrax  bacillus  in  special  culture  fluids 
gave  rise  to  a  substance  which,  when  injected  into  the  organism, 
protected  not  only  against  an  immediate  but  also  subsequent  attacks. 

In  1889  Hankiri  worked  under  the  guidance  of  Koch  in  the 
Hygienic  Institute  of  Berlin.  The  acquired  tolerance  of  the  effect 
of  ordinary  albumoses,  and  the  experiments  of  Sewall,  who  pro- 
duced immunity  against  lethal  doses  of  the  albumose  of  snake 
poison  by  the  injection  of  minute  doses,  led  Hankin  to  expect  that 
an  albumose  developed  in  anthrax  cultures,  and  that  the  anthrax 
albumose  would  probably  confer  immunity  from  the  disease.  Haiikin. 
succeeded  in  isolating  it  from  culture  fluids.  It  was  precipitated  by 
excess  of  absolute  alcohol,  well  washed  in  alcohol  to  free  it  from 
addition  of  ptomaines,  filtered,  dried,  then  redissolved  and  filtered 
through  a  Chamberland  filter.  With  this  substance  Hankin  suc- 
ceeded in  producing  immunity  in  mice  and  rabbits. 

Sidney  Martin,  working  quite  independently,  grew  anthrax  bacilli 
in  a  solution  of  pure  alkali  albumin  made  from  serum  proteids.  After 
ten  or  fifteen  days  the  organisms  were  removed  by  filtration  through 
a  Chamberland  filter.  The  filtrate  contained  pvoto- albumose  and 
deutero- albumose,  a  trace  of  peptone,  an  alkaloid,  and  small  quantities 
of  leucin  and  tyrosin.  The  mixture  of  albumoses  proved  poisonous 
to  mice.  The  anthrax  alkaloid  produced  symptoms  and  lesions 
similar  to  the  albumoses,  but  much  more  rapidly  and  severely.  It 
is  an  amorphous  yellow  body,  soluble  in  alcohol  and  alkaline  in 
reaction.  Martin  concluded  that  the  anthrax  bacillus  formed  the 
albumoses  and  the  alkaloid  by  digesting  the  alkali  albumin ;  and 
suggested  that  the  alkalinity  of  the  albumoses  explained  their  toxic 


CHEMICAL  PRODUCTS  OF  BACTERIA.  43 

properties,  the  alkaloid  probably  being  in  a  nascent  condition  in  the 
albumose  molecule. 

Tuberculosis.  —Koch  prepared  a  glycerine  extract  of  the 
product  of  the  tubercle  bacillus  in  pure  cultivations,  and  found 
that  the  injection  of  small  doses  produced  a  remarkable  reaction, 
both  local  and  general,  in  tubercular  cases,  and  especially  lupus. 
This  extract,  called  tuberculin,  came  to  be  extensively  used  as  a 
therapeutic  agent,  but  with  disappointing  results.  Very  shortly 
after  the  first  announcement  of  Koch's  discovery,  the  author,  in 
conjunction  with  Herroun,  investigated  the  chemical  properties  and 
physiological  effects  of  the  products  of  the  tubercle  bacillus. 
Cultures  in  glycerine-broth  were  filtered  through  porcelain,  and 
a  clear  amber-coloured  liquid  was  obtained,  which  gave  important 
and  suggestive  chemical  reactions.  As  this  filtrate  contained  the 
products  of  the  growth  of  the  bacillus  most  probably  in  minute 
quantities,  it  was  evaporated  at  a  low  temperature  over  sulphuric 
iicid.  The  viscous  residue  was  dissolved  in  distilled  water  and 
tested  on  the  healthy  guinea-pig.  The  result  was  a  marked  fall  of 
temperature,  staring  coat,  extreme  irregularity  of  the  heart's  action, 
muscular  spasms,  loss  of  control  over  the  extremities,  and  death. 

A  preliminary  examination  of  glycerine -broth  cultivations  having 
.shown  the  presence  of  non-coagulable  proteid  bodies  of  the  nature 
of  albumose  and  peptone,  and  a  crystallisable  precipitate  of  a 
remarkable  character  resulting  on  the  addition  of  iodine,  the  idea, 
naturally  suggested  itself  that  the  tubercle  bacillus  might  form 
albumoses  and  an  alkaloid  or  ptomaine  similar  to  the  substances 
isolated  by  Martin  from  pure  cultivations  of  the  Bacillus  anthraois. 

Koch  pointed  out  that  the  effective  substance  in  his  extract 
could  be  precipitated  by  absolute  alcohol ;  the  author  and  Herroun 
determined  to  investigate  the  properties  and  physiological  effects 
of  the  separated  products.  They  accordingly  set  to  work  to  isolate 
the  ptomaine,  of  the  existence  of  which  they  had  some  qualitative 
indication,  and  at  the  same  time  to  examine  the  properties  of  the 
albuminous  bodies. 

In  this  endeavour  the  general  method  they  found  satisfactory  was 
as  follows.  The  clear  filtrate  from  the  culture  was  evaporated  at 
40°  C.  to  a  very  small  bulk,  and  the  residue  thus  obtained  was 
mixed  with  an  excess  of  absolute  alcohol,  which  precipitated  the 
albumoses  and  peptone.  It  was  found  that  by  adding  the  alcohol 
by  degrees  a  partial  separation  of  the  albuinose  from  the  peptone 
could  be  effected,  the  latter  being  only  precipitated  when  the  alcohol 
was  nearly  absolute.  The  precipitated  albumose  was  collected  on 


44  BACTERIOLOGY. 

a  filter  and  redissolved  in  distilled  water.  In  another  experiment 
the  albumose  underwent  a  second  precipitation,  and  after  washing 
was  again  dissolved. 

The  alcoholic  filtrate  from  the  precipitated  albuminous  bodies  was 
then  concentrated  at  a  very  gentle  heat  until  a  viscous  residue  was 
left  containing  the  glycerine  originally  present  in  the  cultivating 
medium  and  the  extractives  and  products  of  the  bacillus  soluble 
in  alcohol.  With  this  residue  definite  reactions  of  an  alkaloidal 
substance  or  ptomaine  were  obtained. 

Careful  experiments,  however,  led  to  the  belief  that  the  whole 
of  the  ptomaine  was  not  separated  from  the  albuminous  precipitate 
by  simple  addition  of  alcohol,  and  the  above  method  was  therefore 
slightly  modified. 

The  ptomaine  is  soluble  in  water  and  alcohol,  and  sparingly 
soluble  in  amyl-alcohol,  but  insoluble  in  benzine,  ether,  or  chloro- 
form, which  liquids  therefore  fail  to  extract  it  from  aqueous 
solutions.  In  its  aqueous  solutions  it  is  distinctly  but  not  strongly 
alkaline  to  test-paper.  Phospho-tungstic  acid  gives  with  it  a  white 
flocculent  precipitate.  Phospho-molybdic  acid  gives  a  pale  yellow 
precipitate,  soluble  in  ammonia  to  a  blue  solution  which  becomes 
colourless  on  boiling.  In  this  respect  it  resembles  the  vegetable  alka- 
loids, aconitiri  and  atropin.  It  must  be  remembered,  however,  that 
albuminous  bodies  are  -precipitated  by  both  this  and  the  preceding 
reagents,  and  in  the  case  of  the  former  a  reduction  of  the  phospho- 
molybdate  giving  the  blue  solution  with  ammonia  is  obtained. 

The  reducing  power  of  the  ptomaine  is  shown  by  the  conversion 
after  a  short  time  of  ferri-cyanide  of  potassium  to  ferro-cyanider 
giving  the  Prussian  blue  test  with  ferric  chloride,  to  which  much 
undue  importance  was  attached  by  Brouardel  and  Boutmy.  The 
solution  of  albumose  and  solution  of  peptone  are  both  capable  of 
giving  this  reaction  as  well  as  many  vegetable  alkaloids.  A  solution 
of  the  ptomaine  is  not  precipitated  by  ferro- cyanide  of  potassium  or 
potassic  bichromate. 

In  strong  solutions  it  yields  precipitates  with  platinic  chloride 
(yellow),  gold  chloride  (pale  yellow),  and  mercuric  chloride  (white). 
That  yielded  by  the  first  of  these  reagents  is  granular  in  character, 
and  quite  insoluble  in  alcohol,  though  apparently  soluble  in  water. 
The  precipitation  by  gold  chloride  excludes  amides  and  ammonium 
salts. 

With  iodine  in  hydriodic  acid  or  potassic  iodide  a  precipitate  is 
obtained  which  is  occasionally  crystalline,  more  often  granular  or 
amorphous. 


CHEMICAL    PRODUCTS    OF   BACTERIA.  45 

This  precipitate  is  soluble  in  alcohol,  and  is  redeposited  when  the 
alcohol  is  evaporated.  On  heating  it  is  redissolved  into  oily  drops  of 
a  dark  colour.  With  picric  acid  a  granular  precipitate  is  obtained, 
which  under  the  microscope  is  seen  to  consist  of  minute  crystals. 
This  precipitate,  on  standing,  is  converted  into  rounded  crystalline 
masses  with  numerous  small  crystals  admixed. 

The  ptomaine  appears  to  be  easily  broken  up  by  heating, 
especially  in  the  presence  of  mineral  acids  or  of  baryta.  The  actual 
quantity  obtained  from  a  considerable  amount  of  culture  fluid  was 
very  small,  and  as  it  was  possible  that  when  the  bacilli  were  grown 
in  a  medium  richer  in  albumin,  such  as  the  animal  body,  more  of 
these  products  might  be  formed,  the  liquid  obtained  by  extracting 
large  masses  of  tubercular  growths  from  cattle  was  examined  in  a 
similar  manner.  In  this  extract,  after  nitration  through  porce- 
lain, an  albumose,  and  minute  quantities  of  a  ptomaine  were 
obtained  which  in  reactions  was  identical  with  that  obtained  from 
the  artificial  cultivation  of  the  bacillus,  but  present  in  even  smaller 
amount.  The  probable  explanation  of  this  is,  that  in  the  living 
animal  the  ptomaine  is  constantly  being  removed  ;  or  it  may  indicate 
that  it  is  only  formed  in  minute  quantity  under  those  conditions. 

Having  succeeded  in  obtaining  the  albumose  and  the  ptomaine 
in  separate  solutions,  we  next  proceeded  to  ascertain  the  effects  of 
these  substances  upon  healthy  and  tubercular  guinea-pigs. 

The  effect  of  the  ptomaine  isolated  from  different  series  of 
cultures  was  as  follows.  A  rise  of  temperature  occurred  in  tuber- 
cular animals,  and  distinct  enlargement  of  tubercular  glands.  There 
was  a  slight  indication  of  a  depression*  of  temperature  or  hypothermic 
effect  on  healthy  animals.  The  albumose,  whether  obtained  from 
pure  cultivations  of  the  bacillus  or  from  tubercular  tissue,  pro- 
duced a  marked  rise  of  temperature  in  tubercular  guinea-pigs.  On 
the  other  hand,  in  a  control  experiment  on  a  healthy  guinea-pig 
there  was  an  equally  well-marked  fall  of  temperature.  The  effect 
upon  the  tubercular  glands  in  the  cases  associated  with  marked  ris3 
of  temperature  was  to  render  them  well-defined,  indurated,  and 
painful,  rather  than  any  considerable  increase  in  volume. 

Hunter  made  a  chemical  examination  of  Koch's  crude  extract, 
and  confirmed  the  presence  of  albumoses  and  alkaloidal  substances. 
The  albumoses  consisted  chiefly  of  proto-albumose  and  deutero-albu- 
mose  with  hetero-albumose,  and  occasionally  a  trace  of  dys-albumose. 
Two  alkaloidal  substances  were  obtained  in  the  form  of  platinum 
compounds  of  their  hydrochlorate  salts.  In  addition  there  were 
extractives,  mucin,  inorganic  salts,  glycerine,  and  colouring-matter. 


OF  THF 

UNIVERSITY 


46  BACTERIOLOGY. 

Swine  Fever. — Schweinitz  applied  Brieger's  methods  in  the 
investigation  of  the  products  of  the  swine  fever,  or  hog  cholera 
bacillus.  Broth- cultures  were  neutralised  with  dilute  hydrochloric 
acid,  and  evaporated  in  the  water  bath.  The  residue  was  treated 
with  96  per  cent,  alcohol,  and  the  filtered  solution  with  mercuric 
chloride.  A  heavy  crystalline  precipitate  was  separated  by  filtra- 
tion, treated  with  water,  and  decomposed  with  sulphuretted  hydrogen, 
and  cadaverin  and  methylamine  were  isolated.  The  filtrate  from 
the  mercuric  chloride  precipitate  was  freed  from  excess  of  mercury 
by  sulphuretted  hydrogen,  and  the  mercury  sulphide  filtered  off. 
The  residue,  after  concentration  of  the  filtrate,  was  extracted  with 
absolute  alcohol,  and  the  solution  showed  the  presence  of  an  alka- 
loidal  salt.  The  double  salt  obtained  with  platinum  chloride  was 
submitted,  after  crystallisation,  to  an  analysis,  and  the  results  gave 
the  formula  (C14H34N2PtCl6).  The  hydro- chloride  is  soluble  in  abso- 
lute alcohol  as  well  as  in  water,  and  produces  needle-like  crystals. 

On  treating  the  culture  fluids  with  excess  of  absolute  alcohol  a 
white  flocculent  precipitate  was  obtained  partly  soluble  in  water,  and 
re- precipitated  by  alcohol.  It  was  obtained  in  the  form  of  white 
crystalline  plates.  A  watery  solution  gives  almost  insoluble  needle- 
crystals  on  the  addition  of  platinum  chloride.  These  products  were 
respectively  termed  sucholo-toxin  and  sucholo-albumin.  Small  doses 
of  these  substances  produce  in  guinea-pigs  a  slight  rise  in  tempera- 
ture, and  ulcaration  at  the  seat  of  injection.  Large  doses  produce 
a  fatal  result  in  six  to  twenty-four  hours.  Schweinitz  asserts  that 
he  has  produced  immunity  in  guinea-pigs.  An  attempt  to  produce 
immunity  in  swine  by  injection  of  the  albumose  gave  unsatisfactory 
results. 

Diphtheria. — Roux  and  Yersin  finding  that  filtered  cultures  of 
the  diphtheria  bacillus  produced  paralysis,  affecting  chiefly  the  hind 
legs,  and  a  fatal  result  in  rabbits  and  guinea-pigs,  proceeded  to 
investigate  the  chemical  products.  They  succeeded  in  obtaining  a 
white  amorphous  substance  which  was  extremely  active  when 
injected  into  guinea-pigs.  It  was  precipitated  by  alcohol  from  an 
aqueous  solution,  and  it  was  calculated  that  '0004  gram  would 
destroy  eight  guinea-pigs  of  400  grams,  or  two  rabbits  of  3  kilos, 
each.  They  concluded  that  the  poison  was  an  enzyme  or  ferment, 
as  it  not  only  acted  in  extremely  small  doses,  but  it  was  attenuated 
by  heat  and  destroyed  by  boiling. 

Brieger  and  Frankel  confirmed  these  experiments,  and  asserted 
that  the  poison  was  a  tox-albumin ;  but  according  to  Martin  their 
chemical  analysis  and  reactions  were  vitiated  by  the  fact  that  they 


<  HKMICAL    PRODUCTS   OF   BACTERIA.  47 

had  peptone  in  their  cultivating  medium.  Martin  examined  the 
products  by  using  as  a  culture  medium  a  1  to  2  per  cent,  solution 
of  alkali-albumin  in  broth  made  from  beef,  omitting  the  peptone. 
After  about  thirty  days  the  bacillus  had  converted  the  alkali-albumin 
into  albumoses,  which  gave  the  reactions  of  proto-  and  deutero- 
albumose,  with  small  quantities  of  an  organic  acid.  A  single  dose 
of  these  albumoses  produced  weakness  of  the  hind  limbs,  which  after 
a  time  passed  off.  The  animal  was  killed,  and  the  nerves  which 
were  examined  showed  degeneration.  Repeated  intravenous  in- 
jection on  successive  days,  amounting  in  all  to  a  dose  of  1'69  grams 
per  kilo,  of  body  weight,  produced  high  fever,  followed  by  depres- 
sion of  temperature,  severe  watery  diarrhoea,  and  emaciation.  The 
taudon  reflexes  began  to  diminish  after  the  ninth  day,  on  the 
eleventh  or  twelfth  day  there  was  definite  paralysis  of  the  hind  legs, 
and  on  the  seventeenth  day  reflexes  could  scarcely  be  obtained. 

Martin  thus  gives  his  method  of  abstracting  the  poisonous  pro- 
ducts either  from  cultures  or  from  diphtheritic  tissues.  In  dealing 
with  tissues,  the  spleen  and  other  organs  are  first  finely  minced  and 
placed  in  rectified  spirit,  and  the  blood  is  also  placed  in  spirit,  and 
allowed  to  stand  till  the  proteids  are  coagulated ;  they  are  then 
filtered,  and  the  residue  extracted  with  cold  water,  all  the  extracts 
are  mixed  together,  and  evaporated  at  35°  C.  to  a  small  bulk,  and 
thrown  into  absolute  alcohol.  Most  of  the  albumoses  are  precipi- 
tated, the  alcohol  is  poured  off,  evaporated  to  dryness  at  a  low 
temperature,  and  extracted  by  absolute  alcohol  until  nothing  more 
dissolves.  The  residue  is  deutero- album ose  and  mineral  salts.  All 
the  proteid  is  mixed  together,  dissolved  in  water,  and  precipitated  by 
alcohol,  the  process  being  repeated  to  remove  any  traces  of  bodies 
soluble  in  alcohol  and  the  excess  of  mineral  salts.  At  the  last 
precipitation  the  precipitate  is  allowed  to  stand  under  alcohol  for 
about  two  months.  The  alcohol  is  then  poured  off,  and  the  pre- 
cipitate dried  in  vacuo. 

The  resulting  product  is  a  light  yellowish -brown  powder  soluble 
in  water,  cold  or  boiling,  giving  a  yellowish  and  faintly  acid  or 
nearly  neutral  reaction.  It  is  composed  of  deutero-albumose  with 
a  slight  amount  of  proto-albumose  but  no  peptone.  It  gives  the 
ordinary  actions  of  proteids  and  a  well-marked  biuret  reaction.  It 
is  precipitated  from  solution  by  ammonium  sulphate,  and  slightly 
by  nitric  acid.  The  reactions  are  similar  to  those  of  peptic  deutero- 
allmmose.  The  alcoholic  extract  of  the  tissues  is  strongly  acid,  and 
contains  free  fatty  acid  and  an  organic  acid  insoluble  in  chloroform. 
The  organic  acid  is  readily  soluble  in  water  and  absolute  alcohol, 


48  BACTERIOLOGY. 

and  insoluble  in  ether,  chloroform,  and  benzine.  It  is  a  yellowish 
amorphous  body,  becoming  a  deep  brown  when  made  alkaline. 

Martin  concludes  that  whereas  the  Bacillus  anthracis  produces 
albumoses  and  an  organic  base,  in  diphtheria  we  find  albumoses  and 
an  organic  acid. 

Glanders.— Kalming  has  obtained  from  cultures  of  the  glanders 
bacillus  an  extract  similar  to  tuberculin.  This  crude  extract  is 
known  as  mullein,  and  is  extensively  used  for  the  diagnosis  of 
glanders.  In  a  glandered  horse  it  causes  a  rise  of  temperature 
and  swelling  at  the  seat  of  the  injection,  and  the  glandered  nodules 
become  swollen  and  painful.  Finger  claims  to  have  produced 
immunity  from  glanders  by  inoculation  of  the  products  contained 
in  sterilised  cultures.  Schweinitz^  extracted  from  cultures  a  non- 
poisonous  albumose,  and  obtained  only  traces  of  a  ptomaine. 

Suppuration  and  Pneumonia. — Brieger  obtained  a  ptomaine 
from  cultures  of  Staphylococcus  pyogenes  aureus,  and  Roux  and 
Yersin  a  tox-albumiri  fatal  to  rabbits  and  guinea-pigs  in  a  few 
days.  There  was  pus-formation  at  the  seat  of  inoculation,  with 
redness  and  swelling  of  the  surrounding  parts. 

From  pure-cultures  of  the  micrococcus  of  pneumonia  Klemperer 
obtained  a  tox-albumin,  for  which  the  name  pneumo-toxin  has  been 
suggested. 

ENZYMES  OR  FERMENTS. 

Many  bacteria  liquefy  the  nutrient  gelatine  in  which  they  are 
cultivated.  This  is  due  to  the  development  of  a  ferment  or  enzyme, 
which  dissolves  the  albumin  and  gelatine. 

Enzymes  are  products  of  the  vital  activity  of  living  bacteria.. 
Bitter,  and  independently  Sternberg,  showed  that  when  a  liquefying 
bacterium  is  removed  by  filtration  or  destroyed  by  heat,  the  culture 
fluid  retains  the  power  of  liquefying  gelatine.  As  this  occurs 
usually  when  the  reaction  is  alkaline,  bacterial  enzymes  resemble 
trypsin  and  papain  rather  than  pepsin.  They  can  be  extracted  with 
glycerine,  and  are  quite  harmless.  If  injected  into  animals  no  effect 
is  produced,  and  after  a  few  hours  no  trace  of  them  can  be  found. 
According  to  Fermi,  the  influence  of  temperature  on  the  enzymes 
produced  by  different  bacteria  will  be  found  to  vary  very  consider- 
ably. The  enzyme  of  Staphylococcus  pyogenes  aureus  is  destroyed 
at  55°  C.,  while  the  enzyme  of  Bacillus  anthracis  succumbs  at  a 
temperature  of  65°  C.  to  70°  C. 

Some  bacteria  produce  both  enzymes  and  toxins,  but  many  pro- 
duce enzymes  and  not  toxins,  and  others  toxins  but  not  enzymes. 


CHAPTER    V. 

IMMUNITY. 

THE  condition  of  being  insusceptible  to  an  infective  disease  may  be 
either  natural  or  acquired.  In  studying  the  pathogenic  organisms 
several  examples  of  natural  immunity  will  be  encountered.  The 
bacillus  of  septicaemia,  so  fatal  to  house  mice,  has  been  shown  to 
have  no  effect  upon  field  mice.  The  bacillus  of  anthrax  is  innocuous 
to  cats  and  white  rats.  The  bacterium  of  rabbit  septicaemia  is 
equally  inert  in  dogs,  rats,  and  guinea-pigs.  The  immunity  may 
be  as  in  these  cases  complete,  or  only  partial.  .  Ordinary  sheep  are 
very  easily  affected  with  anthrax,  but  Algerian  sheep  succumb  only 
to  large  doses  of  the  virus.  Natural  immunity  may  not  only  be 
characteristic  of  certain  species,  but  it  may  occur  in  certain  indi- 
viduals of  a  susceptible  species.  The  same  immunity  occurs  in  man, 
for  certain  individuals,  though  equally  exposed  during  an  epidemic 
of  small-pox,  may  escape,  whereas  others  readily  fall  victims  to  the 
disease. 

Acquired  immunity  is  illustrated  by  the  protection  afforded  by 
one  attack  of  the  exanthemata  against  subsequent  attacks.  Thus 
one  attack  of  measles  or  small-pox,  as  a  rule,  affords  complete 
protection.  A  knowledge  of  the  immunity  resulting  in  the  latter 
case  led  to  the  introduction  of  inoculation  of  small- pox  as  a 
protection  against  natural  small-pox. 

Immunity  may  be  acquired  by  acclimatization,  for  the  inhabit- 
ants of  tropical  climates  are  less  susceptible  to  the  diseases  of  the 
country,  malarial  fevers,  for  instance,  than  strangers. 

In  civilised  communities  also,  there  appears  to  be  a  degree  of 
acquired  immunity,  for  infectious  diseases  like  measles  introduced 
among  savages  or  isolated  communities  have  assumed  the  most 
malignant  type. 

The  immunity  acquired  by  protective  inoculation  constitutes,  in 
connection  with  the  study  of  pathogenic  micro-organisms,  a  subject 
of  pre-eminent  interest  and  importance.  Pasteur,  in  his  researches 

49  4 


50  BACTERIOLOGY. 

upon  fowl-cholera,  observed  that  after  non-fatal  cases  the  disease 
either  did  not  recur,  or  the  severity  of  a  subsequent  attack  was  in 
inverse  proportion  to  the  severity  of  the  first  attack.  It  occurred 
to  him  to  endeavour  to  obtain  the  virus  of  this  disease  in  a  form 
which  would  provoke  a  mild  attack  of  the  disease,  and  thus  give 
protection  against  the  virulent  form.  This  attenuation  or  miti- 
gation of  the  virus  was  successfully  attained  by  allowing  cultiva- 
tions of  the  microbe  in  chicken-broth  to  remain  with  a  lapse 
of  several  months  between  the  carrying  on  of  successive  cultiva- 
tions in  fresh  media.  The  new  generations  which  were  then 
obtained  were  found  to  have  diminished  in  virulence,  and  ultimately 
a  virus  was  obtained  which  produced  only  a  slight  disorder ;  on 
recovery  the  animal  was  found  to  be  proof  against  inoculation  with 
virulent  matter.  The  explanation  given  by  Pasteur  of  this  change 
was,  that  prolonged  contact  with  the  oxygen  of  the  air  was  the 
influence  which  diminished  the  virulence,  and  he  endeavoured  to 
prove  this  by  showing  that  when  broth  was  inoculated  in  tubes 
which  could  be  sealed  up,  so  that  only  a  small  quantity  of  air 
was  accessible  to  the  microbe,  the  virulence  of  the  cultures  was 
retained. 

Toussaint  investigated  the  possibility  of  attenuating  the  virus  of 
anthrax.  Sheep  injected  with  3  cc.  of  defibrinated  blood,  con- 
taining anthrax  bacilli,  which  had  been  exposed  to  55°  C.  for  ten 
minutes,  recovered,  and  were  afterwards  insusceptible.  Pasteur 
subsequently  argued  that  this  method  did  not  admit  of  practical 
application,  because  difficulties  would  arise  in  dealing  with  infective 
blood  in  quantity,  and  artificial  cultivations  started  from  this  blood 
could  not  be  relied  upon,  as  they  proved  sometimes  as  virulent  as 
ever. 

Pasteur  endeavoured  to  apply  the  same  method  for  obtaining 
an  attenuated  virus  of  anthrax,  as  he  had  successfully  employed 
in  fowl-cholera.  A  difficulty  was  soon  encountered,  for  in  culti- 
vations of  this  bacillus,  with  free  access  of  air,  spore-formation 
readily  takes  place,  and  the  spores  are  well  known  to  have  an 
extraordinary  power  of  retaining  their  virulence.  Pasteur  found 
that  the  bacilli  ceased  to  develop  at  45°  C.,  and  he  believed  that 
spore-formation  ceased  at  42°  to  43°  C.,  the  bacilli  continuing  to 
develop  by  fission  only.  The  cultivations  were,  therefore,  kept  at 
this  temperature,  and  at  the  end  of  eight  days  the  bacilli  were 
found  to  have  lost  their  virulence,  and  were  quite  inert  when 
inoculated  in  guinea-pigs,  sheep,  or  rabbits.  This  total  destruction 
was,  however,  preceded  by  a  gradual  mitigation,  so  that  a  virus 


IMMUNITY.  51 

could  be  obtained,  by  taking  it  at  the  right  time,  which  gave  only  a 
mild  disease,  and  afforded  subsequent  protection. 

At  Melun,  in  1881,  the  protective  inoculation  against  anthrax 
was  put  to  a  practical  test.  Sheep  and  oxen  were  inoculated  with 
the  mitigated  virus,  and  then  with  a  virulent  form ;  at  the  same 
time  other  sheep  and  oxen  were  inoculated  with  the  virulent  form 
without  previous  vaccination,  as  a  control  experiment.  The  unpro- 
tected sheep  died  without  exception ;  the  unprotected  oxen  suffered 
from  oedematous  swellings  at  the  seat  of  inoculation,  and  a  rise  of 
temperature  ;  but  all  the  protected  animals  remained  healthy. 

As  a  result  of  these  experiments  an  idea  arose  that  by  preventive 
inoculation  with  attenuated  virus  all  communicable  diseases  would 
in  time  be  eradicated ;  but  this  does  not  follow,  for  all  communi- 
cable diseases  do  not  confer  immunity  after  a  first  attack ;  in 
influenza  the  very  reverse  is  believed  to  occur,  and  erysipelas  of  the 
face  leads  to  an  increased  liability  to  subsequent  attacks.  Even 
with  regard  to  the  prevention  of  anthrax,  Pasteur's  researches  were 
opposed  and  criticised.  Koch  investigated  the  subject,  and  came  to 
the  conclusion  that  the  process  did  not  admit  of  practical  applica- 
tion, chiefly  on  the  ground  that  as  immunity  lasted  only  a  year,  the 
losses  from  the  vaccination  process  would  be  as  great  or  even 
greater  than  from  the  spontaneous  disease ;  further,  there  was 
danger  in  disseminating  a  vaccine  of  the  strength  required  to  be 
effectual. 

Chauveau  proved  that  the  attenuation  was  due  to  the  tempera- 
ture, and  not  to  the  prolonged  effect  of  oxygen.  By  keeping 
cultivations  at  42°  to  43°  C.  in  vacua,  the  virulence  was  found 
to  disappear  in  twenty-four  hours,  and  by  keeping  cultivations 
at  a  low  temperature  with  free  access  of  air,  the  virulence  was 
retained.  Chauveau  considered,  therefore,  not  only  that  oxygen  was 
not  the  agent,  but  that  the  mitigation  was  much  more  easily  effected 
in  its  absence.  In  spite  of  these  adverse  criticisms,  these  researches 
nevertheless  confirmed  the  principle  of  Pasteur's  conclusion,  that 
immunity  could  be  induced  by  experimental  measures,  and  further 
showed  that  he  had  considerably  advanced  the  methods  by  which  this 
could  be  effected. 

Chauveau  succeeded  also  in  attenuating  the  virus  by  a  modifica- 
tion of  Toussaint's  method.  Sterilised  broth  was  inoculated  with 
the  bacilli,  and  placed  in  the  incubator  at  42°  to  43°  C.  After  the 
lapse  of  twenty  hours  it  was  removed  to  another  incubator  at  47°  C. 
According  to  the  time  of  exposure  to  this  increased  temperature,  the 
mitigation  varied  in  degree.  Thus  inoculation  with  the  virus,  before 


52  BACTERIOLOGY. 

it  was  exposed  to  47°  C.,  was  fatal  to  guinea-pigs;  but  after  one 
hour  at  47°  C.  the  virulence  was  diminished,  and,  though  ultimately 
fatal,  life  was  prolonged ;  after  two  hours'  exposure  at  47°  C.  only 
half  the  animals  died ;  and  after  three  hours'  exposure  they 
recovered,  and  were  rendered  refractory  to  subsequent  inoculation. 

Attenuation  of  the  virus  of  anthrax  has  also  been  induced  by 
chemical  means.  Chamberland  and  Roux  stated  that  a  fresh  growth 
started  from  a  cultivation  of  bacilli  which  had  been  subjected  for 
twenty- nine  days  to  F^  of  carbolic  acid  was  found  to  be  inert  in 
guinea-pigs  and  rabbits.  Bichromate  of  potash  added  to  a  cultiva- 
tion in  the  proportion  of  y^J^-g-  to  -g^f  gave,  after  three  days,  a 
new  growth,  which  killed  rabbits,  guinea-pigs,  and  half  the  sheep 
inoculated  •  after  ten  days,  rabbits  and  guinea-pigs,  but  not  sheep ; 
and  after  a  longer  time  even  guinea-pigs  were  unaffected. 

In  other  diseases  similar  results  have  been  obtained.  Arloing, 
Cornevin,  and  Thomas  found  that  by  inoculating  a  small  quantity 
of  the  virus  of  symptomatic  anthrax  anywhere  in  the  subcutaneous 
connective  tissue,  or  a  moderate  quantity  at  the  root  of  the  tail, 
and  even  by  intravenous  injection,  immunity  was  obtained  from  a 
virulent  dose. 

In  swine-erysipelas,  Pasteur  and  Thuillier  obtained  attenuated 
virus  upon  quite  another  principle.  They  discovered  that  by 
passing  the  virus  through  pigeons  the  virulence  was  increased,  but 
by  passing  it  through  rabbits  it  was  progressively  diminished.  Thus 
a  virus  was  obtained  from  the  rabbit,  which  produced  only  a  mild 
disease  in  pigs,  and  after  recovery  complete  immunity.  Similarly 
in  rabies,  Pasteur  found  that  passage  of  the  virus  through  various 
animals  considerably  modified  its  properties.  By  inoculating  a 
monkey  from  a  rabid  dog,  and  then  passing  the  virus  through  other 
monkeys,  the  virulence  was  diminished  ;  but  by  inoculating  a  rabbit 
from  the  dog,  and  passing  the  virus  from  rabbit  to  rabbit,  the 
virulence  increased. 

In  rabies,  Pasteur  has  employed  another  method  of  attenuating 
the  virus.  The  spinal  cord  of  inoculated  rabbits  is  removed  with 
all  possible  precautions,  and  portions  a  few  centimetres  in  length 
are  suspended  in  flasks  in  which  the  air  is  dried  by  fragments  of 
potash.  By  this  process  the  virulence  is  found  to  gradually  diminish 
and  finally  disappear.  Animals  inoculated  with  portions  of  these 
cords,  after  suspension  for  a  certain  time,  are  rendered  refractory  to 
inoculation  with  virulent  cords.  Having  rendered  dogs,  which  had 
been  previously  bitten,  free  from  the  supervention  of  symptoms  of 
hydrophobia  by  means  of  protective  inoculation,  Pasteur  proceeded 


IMMUNITY.  53 

to  apply  the  same  treatment  to  persons  bitten  by  rabid  animals,  with 
results  which  tend  to  the  belief  that  a  real  prophylactic  for  rabies 
has  been  discovered. 

Immunity  may  also  be  produced  by  injecting  the  toxic  products 
existing  in  pure  cultivations  after  removal  of  the  bacilli.  Salmon 
was  the  first  to  produce  immunity  in  this  way,  by  utilising  the  toxic 
products  of  the  bacterium  of  hog-cholera,  which  were  separated  by 
nitration  from  the  living  micro-organisms ;  and  shortly  afterwards 
Wooldridge  demonstrated  that  filtered  anthrax  cultures  contained 
a  substance  which  conferred  immunity.  Behring  and  Kitasato 
produced  immunity  by  mixing  cultures  with  terchloride  of  iodine. 
Yaillard  filtered  the  cultures  through  porcelain,  and  attenuated  the 
products  by  heating  at  different  temperatures. 

Lastly,  in  the  course  of  Behring's  and  Kitasato's  experiments,  it 
was  found  that  the  blood  serum  of  animals  rendered  immune  was 
capable  of  conferring  immunity  on  other  animals.  The  injection 
of  the  toxic  products  of  pathogenic  bacteria  leads  to  the  development 
of  substances  in  the  blood  to  which  the  term  "  antitoxin  "  has  been 
applied.  These  protective  substances  neutralise  or  destroy  the 
injected  poison,  and  blood  serum  which  has  thus  been  rendered 
antitoxic  can  be  utilised  to  confer  immunity  on  other  animals. 

Haffkine's  system  of  vaccination  as  a  protection  against  Asiatic 
cholera  is  supposed  to  be  based  upon  the  principle  of  inducing  the 
formation  of  antitoxins  or  defensive  proteids. 


MECHANISM  OF  IMMUNITY. 

E-aulin  has  shown  that  Aspergillus  niger  develops  a  substance 
which  is  prejudicial  to  its  own  growth,  in  the  absence  of  iron  salts 
in  the  nutrient  soil,  and  Pasteur  suggested  that  in  rabies,  side  by 
side  with  a  living  microbe,  there  is  possibly  some  chemical  product 
or  anti-microbe  which  has,  as  in  Haulm's  experiment,  the  power  of 
arresting  the  growth  of  the  microbe.  If  we  accept  the  theory  of 
arrest  by  some  chemical  product,  we  must  -suppose  that  in  the 
acquired  immunity  afforded  by  one  attack  of  an  infectious  disease 
this  chemical  substance  is  secreted,  and,  remaining  in  the  system, 
opposes  the  onset  of  the  micro-organism  at  a  future  time.  In  the 
natural  immunity  of  certain  species  and  individuals  we  must  suppose 
that  this  chemical  substance  is  normally  present. 

Another  theory  is,  that  the  micro-organisms  assimilate  the 
elements  which  they  require  for  their  nutrition  from  the  blood  and 
tissues,  and  render  the  soil  impoverished  or  otherwise  unsuitable  for 


54  BACTERIOLOGY. 

the  development  of  the  same  species  of  micro-organisms  hereafter ; 
this  condition  may  be  -permanent,  or  the  chemical  constitution  of 
the  tissues  may  be  restored  to  normal,  when  immunity  ceases.  If, 
however,  we  explain  acquired  immunity  by  the  result  of  the  growth 
of  a  previous  invasion  of  micro-organisms,  we  are  still  confronted 
with  the  difficulty  of  explaining  natural  immunity. 

A  third  theory  is  that  the  tissues  are  endowed  with  some 
power  of  vital  resistance  to  the  development  of  micro-organisms, 
similar  to  the  vital  resistance  to  coagulation  of  the  blood,  which  is 
supposed  to  exist  in  the  lining  membrane  of  the  healthy  blood- 
vessel; that  in  some  species  arid  individuals  this  exists  to  a  high 
degree,  and  hence  their  natural  immunity.  But  this  does  not 
explain  how  one  attack  confers  immunity  from  a  subsequent  one. 
One  would  expect  that  the  vital  resistance  would  invariably  be 
lowered  by  a  previous  attack,  and  increased  liability  be  the  constant 
result. 

A  fourth  theory  was  propounded  by  Metchnikoff,  who  maintains 
that  immunity  depends  upon  phagocytosis.  If  anthrax  bacilli  are 
inoculated  in  the  frog,  white  blood-cells,  or  phagocytes,  are  observed 
to  incorporate  and  destroy  them  until  they  entirely  disappear,  and 
the  animal  is  not  affected.  But  if  the  animal,  after  inoculation, 
is  kept  at  a  high  temperature,  the  bacilli  increase  so  rapidly  that 
they  gain  the  upper  hand  over  the  phagocytes,  and  the  animal 
succumbs. 

It  has  also  been  suggested  that  bacteria  may  attract  or  repel 
the  phagocytes,  exercising  either  a  positive  or  a  negative  chemio- 
taxis.  This  power  is  supposed  to  depend  upon  some  special  product 
of  the  bacteria  or  possibly  upon  their  toxins,  as  suggested  by  Roux. 
We  must  suppose  that  the  negative  chemio-taxis  has  become  changed 
to  a  positive  chemio-taxis  in  an  immunised  animal,  so  that  the 
phagocytes,  instead  of  withdrawing  and  leaving  the  bacteria  to 
multiply,  are  readily  drawn  into  the  contest  and  destroy  the 
invaders. 

In  septicaemia  of  mice,  the  white  blood-cells  are  attacked  and 
disintegrated  by  the  bacilli  in  a  remarkable  way.  It  is  difficult, 
however,  to  accept  these  observations  as  affording  a  complete  ex- 
planation of  immunity.  It  is  difficult  to  conceive  that  the  leucocytes 
in  the  blood  and  tissues  in  the  field  mouse  are  differently  constituted 
from  those  in  the  house  mouse,  so  that  they  form  an  effectual 
barrier  to  the  onset  of  bacteria  in  the  one  case,  though  so  readily 
destroyed  in  the  other,  or  that  in  acquired  immunity  the  result  is 
due  to  educating  the  phagocytes  to  respond  to  a  positive  chemio-taxis. 


IMMUNITY.  55 

Phagocytosis  cannot  explain  the  immunity  which  results  from 
the  injection  of  filtered  cultures,  or  of  antitoxins,  but  when 
blood  serum  of  immunised  animals  was  shown  to  possess  antitoxic 
properties,  a  new  explanation  of  immunity  was  at  once  forthcoming. 
In  the  light  of  these  discoveries  immunity,  whether  natural  or 
acquired,  was  regarded  as  due  to  the  accumulation  in  the  blood 
and  tissues  of  substances  which  have  the  property  of  counteracting 
partially  or  entirely  the  products  by  which  pathogenic  bacteria 
produce  their  poisonous  effects.  These  antitoxins,  or  protecting 
proteids,  can  be  obtained  not  only  from  the  blood  but  also  from 
the  spleen  and  the  lymphatic  and  other  glands.  They  result 
from  the  metabolism  of  the  cells  of  the  tissues  of  the  body. 
Phagocytes  in  their  conflict  with  bacteria  may  play  a  small 
part,  but  it  is  more  than  probable  that  immunity  is  altogether 
independent  of  phagocytosis. 


CHAPTER   VI. 

ANTITOXINS   AND   SERUM   THERAPY. 

IT  has  been  clearly  shown  by  the  experiments  of  Fodor  and  Nuttall 
that  some  species  of  bacteria  are  killed  by  a  mixture  with  fresh 
blood.  Fodor  pointed  this  out  in  the  case  of  the  anthrax  bacillus, 
and  Nuttall  confirmed  the  experiments,  and  repeated  them  with 
a  number  of  different  species  of  bacteria. 

Behring  and  Nissen  followed  up  this  line  of  inquiry,  and  found 
that  there  was  a  great  difference  in  the  behaviour  of  freshly  drawn 
blood  to  different  bacteria.  In  some  cases  the  bacteria  were 
destroyed,  in  others  their  growth  was  only  retarded,  and  in  others 
again  they  were  not  affected  at  all.  Bouchard  pointed  out  that 
although  the  normal  blood  serum  of  a  rabbit  may  be  used  for  the 
cultivation  of  Bacillus  pyocyaneus,  the  blood  serum  of  a  rabbit,  which 
has  been  rendered  immune,  will  attenuate  or  entirely  nullify  the 
pathogenic  properties  of  the  bacillus. 

Ogata  and  Jasuhara  obtained  similar  results  by  cultivating 
anthrax  bacilli  in  the  blood  of  immune  animals.  Buchner  demon- 
strated that  this  property  of  fresh  blood  belonged  to  the  serum 
and  not  to  the  cellular  elements,  and  strongly  advocated  the  theory 
that  the  force  opposed  to  invading  bacteria  was  to  be  found  in  the 
serum  rather  than  in  phagocytes. 

Similar  experiments  were  made  with  the  bacteria  of  swine-fever, 
and  Emmerich  and  Mastbaum  discovered  that  the  blood  serum  of 
immune  rabbits  could  be  used  as  a  therapeutic  agent  to  prevent 
the  progress  of  the  disease  in  animals  already  showing  symptoms 
of  infection. 

A  new  light  was  thrown  upon  this  question  by  the  experiments 
of  Behring,  Kitasato,  Tizzoni  and  Cattani,  and  others  in  connection 
with  tetanus  and  diphtheria.  In  these  diseases  the  bacteria  tdo  not 
invade  the  body,  but  the  poisonous  principles  elaborated  at  the 
seat  of  inoculation  are  absorbed  into  the  system  arid  produce 
deleterious  effects.  It  was  obvious  that  attention  must  be  turned 
towards  counteracting  or  destroying  these  poisonous  products. 

56 


ANTITOXINS   AND   SERUM   THERAPY. 


57 


It  was  in  this  direction  that  the  experiments  of  Behring  and 
Kitasato,  in  1890,  proved  to  be  of  profound  importance.  It  was 
shown  that  the  blood  serum  of  a  rabbit  rendered  immune  against 
tetanus  or  diphtheria  had  no  destructive  or  retarding  effect  on  the 
growth  of  the  bacilli,  but  it  possessed  the  power  of  neutralising  the 
poison  developed  by  the  agency  of  the  bacilli.  In  short,  the  serum 
was  shown  to  possess  an  antitoxic  instead  of  a  bactericidal  power. 

Hankin  conceived  the  idea  that  this  property  is  due  to  substances 
of  the  nature  of  defensive  proteids,  and  the  blood  serum  of  the 
naturally  immune  rat  was  found  to  contain  a  proteid  body  with  well- 
marked  alkaline  reaction,  possessing  the  power  of  destroying  anthrax 
bacilli.  Injection  of  this  proteid  into  mice,  together  with  fully 
virulent  anthrax  spores,  prevented  the  development  of  the  disease. 
Young  rats  are  susceptible  to  anthrax,  and,  according  to  Hankin, 
they  can  be  protected  from  anthrax  by  injection  of  the  blood  serum 
of  the  parent.  Tizzoni  and  Cattani  expressed  the  opinion  that  the 
antitoxic  substance  in  the  blood  serum  of  animals  rendered  immune 
against  tetanus  is  a  globulin  to  which  they  gave  the  name  tetanus 
antitoxin.  Buchner  proposed  the  term  alexins  (dAe^w,  I  defend), 
to  signify  these  substances.  Hankin  subdivided  them  into  sozins 
and  phylaxins.  Sozins  are  defensive  proteids  occurring  in  normal 
animals ;  phylaxins  are  only  found  in  animals  artificially  immune  ; 
and  each  of  these  are  sub-classed  by  Hankin  according  to  their  power 
of  attacking  the  bacteria  themselves  or  the  products  they  generate. 


Defensive  proteids 

(Hankin) 
Alexins  (Buchner) 


Sozins : 
Defensive 


proteids 


present  in  the  nor- 
mal animal. 


Phylaxins : 
Defensive 
present 


proteids 
in       the 


animal  after  it  has 
artificially  been 
made  immune. 


fMyco-sozins  : 

Alkaline  globulins  from  rat 
(Hankin),  destroying  an- 
thrax bacillus. 

Toxo-sozins : 

Of  rabbit,  destroying  poison 
of  Vibrio  Metchnikovi 
(Gamaleia). 

Myco-phylaxins : 

Of  rabbit,  destroying  pig 
typhoid  bacillus  (Em- 
merich). 

Toxo-phylaxins : 

Of  rabbit,  etc.,  destroying 
diphtheria  and  tetanus 
poisons  (Behring  and 
Kitasato,  anti-toxin  of 
Tizzoni  and  Cattani). 


Tizzoni  and  Cattani  immunised  dogs  and  other  animals  against 
tetanus,  and  employed  the  antitoxin  as  a  therapeutic  agent.     Its 


58  BACTERIOLOGY. 

active  substance  was  precipitated  by  alcohol.  Behring,  Kitasato,  and 
Schiitz  experimented  with  a  view  to  conferring  immunity  upon 
horses.  The  cultures  were  mixed  with  terchloricle  of  iodine,  and 
injected  at  intervals  of  eight  days,  and  the  antitoxic  power  tested 
on  mice.  By  using  increasingly  virulent  cultures,  the  blood  became 
increasingly  antitoxic. 

Vaillard  filtered  tetanus  cultures  through  porcelain,  and  heated 
the  nitrate  at  gradually  diminishing  temperatures.  The  first  in- 
jections were  made  with  10  cc.,  which  had  been  raised  to  60°  C.  for 
an  hour,  then  a  filtrate  was  used  which  had  been  heated  to  55°  C., 
and  lastly,  a  filtrate  which  had  been  heated  to  50°  C.  The  blood 
became  antitoxic,  and  by  injecting  increasing  quantities  of  virulent 
filtrates  the  antitoxic  power  was  rapidly  intensified,  and  animals 
which  were  injected  with  antitoxin  of  full  strength  possessed 
immunity  many  months  afterwards. 

Roux  and  Vaillard  introduced  another  method.  Virulent  cultures 
were  filtered  through  porcelain,  and  the  filtrate  mixed  with  Gram's 
solution  of  iodine  in  iodide  of  potassium.  To  give  immunity  to  a 
rabbit,  3  cc.  of  toxin  with  1  cc.  of  Gram's  solution  were  injected  on 
the  first  day,  and  increasing  doses  of  toxin  mixed  with  increasing 
doses  of  Gram's  solution  on  the  following  days.  The  same  method 
was  applied  to  horses,  sheep,  and  cattle.  The  antitoxin  was  found 
not  only  in  the  blood,  but  in  the  urine  and  saliva,  and  in  the 
milk  in  cows.  With  cows  and  goats  it  is  necessary  to  proceed 
with  the  utmost  care;  while  horses,  on  the  other  hand,  bear  the 
injections  well,  and  are  therefore  more  suitable  for  this  purpose. 
It  is  also  very  easy  to  obtain  large  quantities  of  blood  from  the 
horse  by  inserting  a  trocar  and  cannula  into  the  jugular  vein. 

Frankel  was  the  first  to  produce  immunity  against  diphtheria 
by  injecting  guinea-pigs  with  toxin  which  had  been  heated  to  70°  C. 
Behring  mixed  the  toxin  with  terchloride  of  iodine,  or  employed  small 
doses  of  pure  toxin.  Horses,  sheep,  goats,  and  dogs  were  rendered 
immune. 

PREPARATION  OF  DIPHTHERIA  ANTITOXIN. 

For  the  preparation  of  diphtheria  antitoxin  Roux  cultivates  the 
diphtheria  bacillus  in  alkaline  broth  with  2  per  cent,  of  peptone,  and 
by  preference,  in  flasks  in  which  the  cultivating  liquid  can  be  exposed 
to  a  current  of  moist  air  at  37°  C.  After  about  three  weeks  the 
culture  is  filtered  through  a  Chamberland  filter,  and  if  tested  on 
a  guinea-pig  it  will  be  found  that  y1^  of  a  cc.  will  kill  an  animal 
weighing  five  hundred  grammes  in  forty-eight  hours.  The  diphtheria 


ANTITOXINS   AND   SERUM   THERAPY.  59 

toxin  immediately  before  the  injection  is  mixed  with  ^  of  its 
volume  of  Gram's  solution.  This  is  used  for  several  weeks,  and 
afterwards  only  pure  toxin  is  injected. 

The  horses  employed  for  this  purpose  are  animals  no  longer  fit 
for  work,  and  it  is  necessary  to  inject  them  first  of  all  with  mullein 
to  be  sure  that  they  are  not  suffering  from  glanders. 

In  a  horse  inoculated  by  Roux,  the  injection  began  with  j  cc. 
of  iodised  toxin,  increased  to  1  cc.  by  the  thirteenth  day,  and  the 
injection  continued  daily.  On  the  seventeenth  day  |  cc.  of  pure 
toxin  was  injected,  and  this  was  increased  by  the  forty-first  day 
to  10  cc. ;  and  on  the  forty-third  day  30  cc.  of  pure  toxin  were 
injected,  caiising  pronounced  oadema.  The  doses  were  still  further 
increased,  until  on  the  eightieth  day  250  cc.  were  injected.  In 
two  months  and  twenty  days  the  horse  had  received  800  cc.  of 
toxin. 

On  the  eighty-seventh  day  the  serum  obtained  had  an  immunising 
power  of  over  50,000.  By  this  is  meant  that  a  guinea-pig  resisted 
inoculation  of  \  cc.  of  virulent  diphtheria  culture  when  injected 
twelve  hours  beforehand  with  serum  in  quantity  equal  to  the  ^^J^j 
part  of  its  body  weight. 

There  are  two  tests  which  can  be  applied  to  the  serum.  First, 
the  antitoxic  serum  added  to  diphtheria  toxin  renders  it  inert ;  andy 
secondly,  if  serum  is  injected  into  a  guinea-pig  and  toxin  injected 
several  hours  afterwards,  no  result  follows. 

Several  ways  have  been  suggested  for  estimating  the  immunising 
power  of  the  serum. 

In  Ehrlich's  system,  the  unit  is  represented  by  "1  cc.  of  anti- 
toxic serum,  which,  added  to  "8  cc.  toxin,  will  neutralise  it  so  that 
the  whole  may  be  injected  subcutaneously  in  a  guinea-pig  without 
producing  O3dema.  The  standard  toxin  is  a  toxin  of  which  *3  cc. 
is  fatal  to  1  kilo,  of  guinea-pig. 

But  the  preventive  power  of  the  serum  is  best  expressed  by  the 
result  of  a  subsequent  injection  of  toxin.  The  immunising  power  is 
estimated  by  the  number  of  grammes  of  guinea-pig  which  can  be 
protected  against  the  minimum  fatal  dose  of  toxin  by  1  cc.  of  anti- 
toxic serum. 

The  antitoxic  serum  can  be  kept  in  sterilised  flasks  in  the  dark, 
with  the  addition  of  a  small  piece  of  camphor,  or  it  may  be  dried 
in  vctcaOj  powdered,  and  thus  supplied  in  a  convenient  form  for  trans- 
port. It  has  merely  to  be  dissolved  in  water  before  use. 

Klein  employed  a  modified  plan  by  which  he  claimed  to  have 
obtained  antitoxin  in  a  far  shorter  time  than  is  possible  by  Roux's 


€0  BACTERIOLOGY. 

method.  Unfiltered  attenuated  cultures  were  injected  into  the 
horse.  Later,  large  quantities  of  living  diphtheria  bacilli  from 
the  surface  of  solid  cultures,  of  gradually  increasing  virulence,  were 
repeatedly  injected  so  as  to  allow  the  bacilli  to  grow  and  multiply. 
In  twenty-three  days  an  antitoxic  serum  was  obtained,  one  part 
of  which  was  found  capable  of  protecting  20,000  to  40,000 
grammes  of  guinea-pig  against  more  than  a  fatal  dose  of  both  living 
bacilli  and  the  resulting  toxin. 

Serum  Treatment  of  Diphtheria. —The  results  obtained  by 
Behring,  Ehrlich,  Kossel,  and  Wasserman,  in  the  treatment  of 
diphtheria  in  children  in  Germany  by  means  of  the  curative  serum, 
and  by  Roux  and  others  in  France,  led  to  the  adoption  of  the  treat- 
ment in  this  country.  It  is  best  to  use  an  especially  constructed 
hypodermic  syringe,  which  can  be  easily  taken  to  pieces,  and  placed 
in  boiling  water  to  sterilise  it.  The  skin  surface  of  the  flank  is 
washed,  and  disinfected  with  1  in  20  carbolic,  and  the  antitoxin  is 
then  injected.  The  syringe  is  taken  to  pieces,  placed  again  in  boiling 
water,  and  thoroughly  cleaned. 

The  dose  will  depend  upon  the  age  of  the  patient  and  the 
strength  of  the  serum.  From  10  cc.  to  20  cc.  are  injected  in  children 
under  fifteen,  and  30  cc.  to  40  cc.  in  older  patients,  and  the 
injection  maybe  repeated  in  12  hours.  The  best  results  are  said 
to  be  obtained  by  injecting  every  12  hours,  for  the  first  12,  36  or 
48  hours,  according  to  the  nature  of  the  case,  1,000  Behring's  units, 
this  being  the  dose  calculated  according  to  the  immunising  power  of 
the  serum.  The  result  of  the  injection  is  to  lower  the  temperature 
and  pulse,  but  frequently  the  reverse  occurs,  and  in  about  half  the 
cases  an  urticarial  and  sometimes  a  scarlatiniform  rash  is  produced. 
Pains  in  the  joints,  in  rare  cases  effusion,  may  also  result  from  the 
injection. 

The  beneficial  results  of  the  treatment  are,  according  to  the 
Report  of  the  Medical  Superintendents  of  the  hospitals  of  the 
Metropolitan  Asylums  Board,  as  follows  : — 

(1)  Diminution  of  the  faucial  swelling  and  of  the  consequent 
distress ; 

(2)  Lessening  or  entire  cessation  of  the  irritating  and  offensive 
discharge  from  the  nose  ; 

(3)  Limitation  of  the  extension  of  membrane ; 

(4)  Earlier  separation  of  the  exudation  ; 

(5)  Limitation  and  earlier  separation  of  membrane  in  laryngeal 
cases ; 

(6)  Improvement  in  general  condition  and  aspect  of  patients  ; 


ANTITOXINS   AND   SERUM   THERAPY.  61 

(7)  Prolongation  of  life,  in  cases  which  terminate  fatally,  to  an 
extent  not  obtained  with  former  methods  of  treatment. 

Statistics  have  also  been  brought  forward  which  show,  assuming 
them  to  be  reliable,  a  great  reduction  in  the  mortality  after  the 
antitoxin  treatment.  A  few  instances  may  be  quoted  to  illustrate 
the  statistical  evidence. 

According  to  Behring,  in  the  four  years  prior  to  the  employment 
of  antitoxin,  there  were  in  Berlin  15,958  cases  of  diphtheria,  with  a 
mortality  of  35*2  per  cent.  In  1894-5  there  was  an  epidemic  of 
5,578  cases.  Behring  asserts  that  if  the  mortality  had  not  been 
reduced  by  the  antitoxin  treatment  1,963  would  have  died  instead 
of  1,056.  Behring  also  states  that  in  the  Charite  Hospital  there 
were  299  patients,  with  53  deaths,  or  16*7  per  cent.  In  the  Bethania 
Hospital,  where  antitoxin  was  not  employed,  there  were  249 
patients,  with  112  deaths,  or  43  per  cent. 

At  Vienna,  at  the  Anna  Hospital  for  children,  the  mortality 
in  760  cases  was  50'65  per  cent.,  but  after  the  introduction  of  anti- 
toxin there  were  40  deaths  in  159  cases,  giving  a  mortality  of 
2 5 '5  per  cent. 

In  New  York,  it  is  said  that  before  the  introduction  of  anti- 
toxin the  mortality  ranged  from  30'67  to  37*34,  while  in  1895,  under 
treatment  with  antitoxin,  the  mortality  fell  to  19*43;  but  it  was 
also  pointed  out  that  since  the  introduction  of  antitoxin  many 
children  with  trifling  attacks  had  been  treated,  and  reported  as 
suffering  from  actual  diphtheria,  and  that  they  would  have  recovered 
without  antitoxin,  and  therefore  these  cases  have  given  the  remedy 
some  credit  which  it  does  not  deserve. 

In  London,  according  to  the  Report  of  the  Medical  Superin- 
tendents of  the  hospitals  of  the  Metropolitan  Asylums  Board  there 
were  in  1894,  before  antitoxin  was  employed,  3,042  cases  of 
diphtheria  with  902  deaths  or  29*6  per  cent.,  and  in  1895,  when 
antitoxin  was  used,  3,529  cases  with  796  deaths  or  22*5  per  cent.  : 
a  reduction  of  7*1  per  cent,  below  that  of  1894.  The  conclusions 
drawn  from  the  statistical  and  clinical  observations  are  summed  up 
in  the  Report  thus  : — 

The  improved  results  in  the  diphtheria  cases  treated  during  the  year 
1895,  are  :— 

(I.)  A  great  reduction  in  the  mortality  of  cases  brought  under  treat- 
ment on  the  first  and  second  day  of  illness. 

(II.)  The  lowering  of  the  combined  general  mortality  to  a  point 
below  that  of  any  former  year. 

(III.)  The  still  more  remarkable  reduction  in  the  mortality  of  the 
laryngeal  cases. 


62  BACTERIOLOGY. 

(IV.)  The  uniform  improvement  in  the  results  of  tracheotomy  at 
each  separate  hospital. 

(V.)  The  beneficial  effect  produced  on  the  clinical  course  of  the 
disease. 

A  consideration  of  the  statistical  tables  and  clinical  observations, 
covering  a  period  of  12  months  and  embracing  a  large  number  of  cases, 
in  our  opinion  sufficiently  demonstrates  the  value  of  antitoxin  in  the 
treatment  of  diphtheria. 

It  must  be  clearly  understood,  however,  that  to  obtain  the  largest 
measure  of  success  with  antitoxin  it  is  essential  that  the  patient  be 
brought  under  its  influence  at  a  comparatively  early  date — if  possible  not 
later  than  the  second  day  of  disease.  From  this  time  onwards  the  chance 
of  a  successful  issue  will  diminish  in  proportion  to  the  length  of  time 
which  has  elapsed  before  treatment  is  commenced.  This,  though 
doubtless  true  of  other  methods,  is  of  still  greater  moment  in  the  case 
of  treatment  by  antitoxin. 

Certain  secondary  effects  not  infrequently  arise  as  a  direct  result  of 
the  injection  of  antitoxin  in  the  form  in  which  it  has  at  present  to  be 
administered,  and,  even  assuming  that  the  incidence  of  the  normal  com- 
plications of  diphtheria  is  greater  than  can  be  accounted  for  by  the 
increased  number  of  recoveries,  we  have  no  hesitation  in  expressing  the 
opinion  that  these  drawbacks  are  insignificant  when  taken  in  conjunction 
with  the  lessened  fatality  which  has  been  associated  with  the  use  of  this 
remedy. 

We  are  further  of  the  opinion  that  in  antitoxin  serum  we  possess 
a  remedy  of  distinctly  greater  valne  in  the  treatment  of  diphtheria  than 
any  other  with  which  we  are  acquainted. 

On  the  other  hand  it  has  been  urged  that  the  decline  in  the 
mortality  in  1895  in  London,  which  has  been  attributed  entirely 
to  the  antitoxin  treatment,  may  possibly  be  partly  due  to  the  pre- 
valence of  a  mild  type  of  the  disease,  and  that  the  fall  in  the 
mortality  during  the  seven  previous  years  from  59  per  cent,  in 
1888  to  29  per  cent,  in  1894,  continued  in  1895. 

It  is  obvious  that  the  whole  subject  requires  to  be  very  carefully 
considered,  and  before  any  final  conclusion  can  be  arrived  at  as  to 
the  therapeutic  value  of  antitoxin,  the  evidence  of  others  who  have 
had  great  experience  in  the  treatment  of  diphtheria  by  the  old  and 
the  new  methods  must  be  taken  into  account,  and  reliable  statistics 
allowed  to  speak  for  themselves. 

PREPARATION  OF  TETANUS  ANTITOXIN. 

Antitoxin  for  use  in  the  serum  treatment  of  tetanus  is  obtained 
from  tlie  horse.  The  tetanus  bacillus  is  cultivated  in  an  atmosphere 
of  hydrogen,  in  flasks  specially  constructed  for  the  purpose.  In 


ANTITOXINS   AND   SERUM   THERAPY.  63 

about  a  fortnight  the  cultures  are  extremely  toxic.  The  toxin  is 
obtained  free  from  bacilli  by  filtration  through  porcelain.  Injec- 
tions may  be  given  daily,  subcutaneously  or  intravenously,  beginning 
with  1  cc.  of  iodised  toxin,  and  gradually  increasing  the  dose  until 
the  pure  toxin  may  be  injected  without  danger. 

Roux  and  Yaillard  produced  immunity  in  about  three  months. 
When  a  few  days  have  elapsed  after  the  last  injection,  the  blood  is 
drawn,  by  means  of  a  trocar  and  cannula,  from  the  jugular  vein  into 
a  sterilised  glass  vessel,  and  set  aside  to  coagulate  ;  next  day  the 
serum  is  drawn  off  with  a  pipette,  and  used  in  the  liquid  state,  or 
dried  in  a  vacuum  over  sulphuric  acid,  and  subsequently  powdered. 
When  required  for  use  the  powder  is  dissolved  in  cold  water.  About 
5  grammes  are  used  for  a  dose. 

Serum  Treatment  of  Tetanus.— The  result,  so  far,  of  the 
employment  of  tetanus  antitoxin  in  animals  suffering  from  tetanus 
is  disappointing,  and  the  serum  treatment  is  not  likely  to  be  of  much 
value  in  veterinary  practice.  Nocard  infected  sheep  with  tetanus 
by  inserting  splinters  of  wood  infected  with  spores  into  the  muscles 
of  the  leg.  Tetanus  supervened  in  eleven  days,  and  the  splinters 
were  removed,  the  tissues  excised,  and  the  wounds  dressed  with 
iodoform.  About  twelve  hours  after  the  symptoms  had  shown 
themselves,  the  sheep  were  inoculated  with  antitoxic  serum  at 
intervals  of  one  hour,  but  they  all  succumbed  to  tetanus.  In  one 
case  the  total  amount  injected  was  160  cc.  of  highly  antitoxic  serum. 

The  antitoxin  has  been  employed  in  tetanus  in  man.  Kanthack 
has  collected  the  history  of  a  number  of  cases,  and  they  indicate 
that  the  treatment  is  useless  in  acute  cases  in  man  with  a  short 
incubation  period,  while  chronic  cases  with  a  long  incubation  period 
often  recover  after  the  treatment.  At  the  same  time  it  must  be 
remembered  that  recovery  often  took  place  in  chronic  cases  before 
the  introduction  of  the  antitoxin  treatment. 

The  question  must  still  be  considered  to  be  sub  jydice,  and  a 
trustworthy  conclusion  can  only  be  based  upon  a  more  extended  use 
of  the  antitoxin  and  impartial  reports  of  every  individual  case. 

ANTITOXIN  OF  SEPTIC  INFECTIONS. 

An  anti-streptococcic  serum  has  been  prepared  by  Marmorek.  A 
culture  of  streptococcus  was  intensified  in  virulence  by  inoculation 
from  rabbit  to  rabbit,  and  highly  virulent  cultures  gave  rise  to  a 
powerful  toxin.  Roget  and  Charrin  also,  found  that  the  serum  of 
immunised  rabbits  and  of  a  horse  conferred  immunity.  A  patient 


OF   THF 

UNIVERSITY 


64  BACTERIOLOGY. 

with  puerperal  fever  was  injected  with  8  cc.,  on  the  follow- 
ing clay  with  16  cc.,  and  on  the  third  day  with  25  cc.  On  the 
fourth  day  the  temperature  had  fallen,  and  the  patient  recovered. 
Favourable  results  are  said  to  have  followed  the  use  of  the  serum 
in  46  cases  of  erysipelas. 

Bokenham,  working  independently,  cultivated  the  streptococcus 
in  a  mixture  of  broth  and  serum.  Horses  and  asses  were  inocu- 
lated, and  a  considerable  degree  of  immunity  established.  The 
serum  of  an  inoculated  ass  possessed  antitoxic  power. 

Ruffer  and  Bullock  succeeded  in  immunising  four  horses  against 
the  toxin  of  Streptococcus  pyogenes  ;  two  had  been  previously  immu- 
nised against  the  toxin  of  the  diphtheria  bacillus.  The  streptococcus 
was  cultivated  by  Marmorek's  methods  in  a  mixture  of  two  parts 
of  blood-serum  and  one  part  of  peptonised  broth,  an'd  the  virulence 
of  cultures  maintained  by  inoculation  of  rabbits.  On  testing  the 
immunising  power  of  the  antitoxic  serum  on  rabbits,  the  effect 
appeared  to  be  slight  in  comparison  with  the  antitoxins  of  the 
bacilli  of  diphtheria  and  tetanus.  In  treating  cases  of  septic  infection 
in  the  human  subject,  it  has  been  recommended  to  commence  with 
two  injections  of  10  cc.,  and.it  is  said  that  no  unfavourable  results 
have  been  met  with  which  could  be  attributed  to  the  effect  of  the 
serum. 

ANTITOXIN  OF  TYPHOID  FEVER  AND  OTHER  DISEASES. 

An  antitoxic  serum  has  been  obtained  by  Chantemesse  for  use  in 
cases  of  typhoid  fever,  and  it  is  probable  that  attempts  will  be  made 
to  extend  the  principle  of  the  antitoxic  treatment  to  other  infective 
diseases. 


CHAPTEK  VII. 

THE    BACTERIOLOGICAL   MICROSCOPE. 

THE  instruments  sometimes  in  use  in  biological  and  pathological 
laboratories  are  not  sufficient  for  the  study  of  bacteria.  It  is 
absolutely  essential  for  the  examination  of  such  minute  objects  that 
the  microscope  should  be  equipped  with  an  objective  of  sufficiently 
high  magnifying  power  and  with  a  special  illuminating  apparatus, 
while  the  mechanical  arrangements  of  the  stage  must  admit  of  the 
examination  of  plate-cultivations.  Tt  would  not  be  within  the  scope 
of  this  work  to  give  a  detailed  account  of  the  mechanical  arrange- 
ments and  optical  principles  of  the  microscope.  These  matters  are 
fully  dealt  with  in  special  works  on  the  subject,*  but  sufficient  will 
be  said  to  afford  assistance  in  the  selection  of  a  suitable  instrument, 
and  to  explain  the  improvements  in  the  microscope  which  have  been 
such  an  aid  in  bacteriological  investigations. 

A  magnified  image  of  an  object  is  the  result  of  the  change 
produced  in  the  direction  of  rays  of  light  which  are  made  to  pass 
through  lenses.  This  alteration  in  the  course  of  the  rays  is  known 
as  refraction.  A  ray  of  light  passing  from  a  larer  into  a  denser 
medium  is  refracted  towards  a  line  drawn  perpendicularly  to  the 
surface  of  the  latter.  A  ray  of  light  passing  through  air  and 
impinging  on  water  will  not  pass  on  in  the  same  direction,  but  will 
be  refracted  towards  a  line  drawn  perpendicularly  towards  the 
surface  of  the  water.  If  the  ray  pass  into  glass  instead  of  water 
a  greater  refraction  will  take  place,  and  if  it  pass  into  diamond  the 
bending  in  its  course  will  be  still  greater  (Fig.  11). 

The  sines  of  the  angle  of  incidence  and  refraction  of  different 
substances  have  a  constant  ratio  to  each  other,  which  is  known 
as  the  index  of  refraction,  and  this  is  determined  for  different 
substances  by  the  refraction  produced  by  the  passage  of  rays  from 
a  vacuum.  Thus  the  index  of  refraction  for  flint  glass  is  about  1*6, 

*  Carpenter  :  The  Microscope,     Nageli  and  Schwenderer :  The  Microscope 
in  Theory  and  Practice. 

65  n 


66 


BACTERIOLOGY 


the  sine  of  the  angle  of  incidence  of  a  ray  passing  from  a  vacuum 
into  glass  being  to  the  sine  of  the  index  of  refraction  as  1*6  to  1. 

If  we  study  the  course  of  a  pencil  of  rays  we  find  that  some 
of  the  rays  are  reflected  instead  of  entering  the  medium  and  being 
refracted.  When,  for  example,  a  pencil  of  rays  falls  upon  water 
or  glass,  after  passing;  through  air,  some  of  the  rays  are  lost  by 
reflection,  and  the  proportion  of  the  lost  rays  will  increase  with 
their  obliquity.  The  diminution  of  the  brightness  of  the  image 
when  pencils  of  rays  have  to  pass  through  lenses  is  thus  accounted 
for,  and  this  loss  of  light  increases  when  the  number  of  surfaces 


FIG.  11. — THE  REFRACTION  OF  LIGHT. 

through  which  the  rays  pass  are,  as  in  high-power  objectives^ 
increased.  There  is  an  additional  loss  when  there  is  an  increase  in 
the  difference  between  the  refractive  power  of  the  different  media 
through  which  light  passes.  When  pencils  of  rays  pass  from  glass 
into  air,  and  then  into  glass  again,  the  loss  is  much  greater  than 
when  the  air  is  replaced  by  a  medium  with  a  refractive  index  more 
nearly  approaching  that  of  glass.  This  explains  the  value  of  the 
immersion  system,  which  will  be  referred  to  more  fully  later  on, 
and  also  the  advantage  of  cementing  pairs  of  lenses  with  Canada 
balsam  or  glass  paste.  The  lenses  used  in  the  optical  arrangements 


THE   BACTERIOLOGICAL   MICROSCOPE. 


67 


of  a  microscope  are  principally  convex,  and  the  imperfections  which 
result  must,  if  possible,  be  entirely  overcome.  These  imperfections 
are  spherical  and  chromatic  aberration. 

Spherical  aberration  results  from  the  unequal  refraction  of 
rays  passing  through  lenses  with  equal  curvatures.  The  rays  passing 
through  an  ordinary  convex  lens  do  not  all  come  to  the  same  focus. 
The  rays  passing  through  the  marginal  portion  come  to  a  focus  at  a 


JPJ 


FIG.  12. — SPHERICAL  ABERRATION. 


point  much  nearer  to  the  lens  than  the  focus  of  the  rays  passing 
through  the  more  central  portion  of  the  lens  (Fig.  1 2)*  If  the  whole 
aperture  of  the  lens  is  used  there  must  of  necessity  be  blurring,  for 
at  the  point  at  which  .the  marginal  rays  form  a  distinct  image  the 
central  rays  will  be  out  of  focus,  and  at  the  point  at  which  the 
central  rays  form  a  distinct  image  the  marginal  rays  will  have 
diverged,  causing  indistinctness. 

This  is  partially  remedied  by  using  a  dia*phragm 
and  shutting  out  the  marginal  rays ;  but  this  is 
at  the  cost  of  loss  of  light  and  diminution  of  the 
angle  of  aperture.  The  difficulty  is  approxiinately 
overcome  in  practice  by  using  a  combination  of 
lenses.  The  aberration  of  a  convex  lens  is  the 
opposite  of  that  of  a  concave  lens  (Fig.  13).  The 
makers  of  the  best  lenses  endeavour  to  obtain  this 
correction  as  perfect  as  possible  to  get  the  sharpness 
of  the  image,  so  essential  in  studying  the  mor- 
phology of  bacteria. 

Chromatic  aberration  is  the  result  of  the 
unequal  refrangibility  of  the  coloured  rays  which 
compose  white  light.  If  parallel  rays  of  light  pass  through  a 
convex  lens  the  violet  rays,  which  are  the  most  refrangible,  will 
come  to  a  focus  at  a  point  much  nearer  to  the  lens  than  the 
focus  of  the  red  rays,  which  are  the  least  refrangible ;  and  the 
intermediate  rays  of  the  spectrum  will  be  focussed  at  points  between 
the  red  and  the  violet.  A  screen  held  at  either  of  these  foci 
will  show  an  image  with  prismatic  fringes  (Fig.  14). 


FIG.  13.— COM- 
BINATION o  F 
LENSES  IN 
ABBE'S  HOMO- 
GENEOUS IM  - 
MERSION. 


68  BACTERIOLOGY. 

The  chromatic  aberration  may  be  reduced  by  stopping  out  the 
marginal  rays;  but  as  it  is  necessary  to  get  the  most  perfect 
correction  possible,  advantage  is  taken  of  the  different  relations 
which  the  refractive  and  dispersive  powers  bear  to  each  other  in 
different  glasses.  By  combining  a  double  convex  lens  of  crown 
glass  with  a  plano-convex  lens  of  flint  glass,  correction  is  obtained 
for  the  violet  and  red  rays.  An  achromatic  objective  is  constructed 
on  this  principle,  but  the  result  is  not  perfect,  as  the  intermediate 
coloured  rays  remain  uncorrected,  and  what  is  termed  a  secondary 
spectrum  gives  rise  to  images  with  coloured  fringes,  especially  at  the 
margin  of  the  field.  Abbe  and  Schott,  after  a  great  number  of 
experiments,  succeeded  in  discovering  a  glass  with  optical  properties 
which  removed  the  secondary  spectrum,  and  objectives  made  with 
the  new  glass  are  termed  apo-chromatic.  There  is  much  more 


FIG.  14. — CHROMATIC  ABERRATION. 

perfect  concentration  of  the  component  rays  than  in  the  ordinary 
achromatic  objectives,  and  the  advantages  thus  obtained  are  very 
great.  The  objectives  can  be  made  of  higher  angle  and  admit  of 
higher  eye-pieces  being  used  without  materially  diminishing  the 
brilliancy  and  definition  of  the  image.  There  is  a  complete  absence 
of  coloured  fringes,  and  the  perfect  definition  is  invaluable  in 
micro- photography. 

Another  fault  which  has  to  be  corrected  is  the  aberration  caused 
by  covering  a  microscopical  preparation  with  a  cover-glass.  Ross 
was  the  first  to  point  out  the  difference  in  the  image  when  the 
object  was  examined  under  a  cover-glass,  and  that  by  altering  the 
position  of  the  front  pair  of  lenses,  in  an  objective  corrected  for  an 
uncovered  object,  the  objective  could  be  corrected  for  the  covered 
object  (Fig.  15). 

Objectives  are  generally  corrected  for  a  standard  thickness  of 
cover-glass,  but  H.  Lister  devised  a  screw-collar  adjustment  by 
which  the  position  of  the  front  pair  of  lenses  could  be  altered  at 
will ;  and  as  it  is  almost  impossible  to  obtain  cover-glasses  which 


THE   BACTERIOLOGICAL   MICROSCOPE.  69 

do  not  vary  slightly  in  thickness,  the  most  perfect  definition 
can  only  be  obtained  by  adjusting  for  each  separate  cover-glass 
preparation. 

Immersion  system. — All  objectives  were  formerly  used  dry — 
that  is  to  say,  with  an  air  space  between  the  objective  and  the 
specimen  to  be  examined — but  high-power  objectives  are  now  almost 
entirely  made  on  the  immersion  system,  a  drop  of  liquid  being 
interposed  between  the  objective  and  the  cover-glass. 

About  fifty  years  ago  Amici  observed  that  if  a  drop  of  water 
intervened  between  the  cover-glass  or  an  uncovered  object  and  the 
lens  the  image  was  more  brilliant.     The  passage  of  rays  from  the 
object  or  the  cover-glass   into   air, 
and  again  from  air  into  glass,  caused 
considerable    loss   of    light.      With 
objectives  of  wide  angle  of  aperture 
the  advantages   were   counteracted 
by  the  reflection  of  rays  falling  ob- 
liquely upon  the  lens.      By  inter-  -,  PPi  I""" ''  mimV3 

posing  water  more  rays  are  bent 
in  or  refracted,  and  enter  the  lens 
instead  of  being  reflected  and  lost. 

Hartnack,    Nachet,   and  others 

adopted  the  immersion  system,  and 

,  .   ,  .  ,  FIG.  15. — OBJECTIVE  WITH  COLLAR 

high-power  water  immersion  lenses  CORRECTION  (b). 

were    constructed  with  high    angle 

of  aperture.*  It  was  found  that  there  was  less  necessity  for 
correcting  for  covers  of  different  thickness,  as  the  aberration  from 
this  cause  was  diminished.  The  lenses  were  corrected  for  an  average 
thickness  of  cover,  and  slight  deviations  produced  hardly  any 
appreciable  effect. 

Wenham,  Stephenson,  Abbe,  and  Zeiss  carried  the  system  to 
perfection.  They  argued  that  the  advantages  obtained  by  water 
immersion  would  be  intensified  if  a  liquid  could  be  found  of  the 
same  refractive  and  dispersive  power  as  crown  glass.  The  media 
would  be  optically  uniform,  and  the  result  a  homogeneous  immersion 
system. 

*  The  angle  of  aperture  is  "  the  angle  made  by  the  most  diverging  of  the 
rays  of  the  pencil  issuing  from  any  point  of  an  object  that  can  enter  the  lens, 
and  take  part  in  the  formation  of  an  image  of  it." 

The  numerical  aperture  is  defined  by  Abbe  as  equal  to  "  the  sine  of  the 
angle  of  aperture  multiplied  by  the  refractive  index  of  the  medium  between 
the  object  and  the  objective." 


70  BACTERIOLOGY. 

After  experimenting  with  different  liquids — solutions  of  salts, 
and  various  essential  oils — Abbe  recommended  cedar  oil  as  most 
suitable  for  the  purpose.  In  its  optical  properties  it  very  closely 
resembles  crown  glass,  and  it  is  far  more  convenient  for  use  than 
any  watery  solutions  of  salts,  especially  when  it  is  necessary  to 
make  a  more  or  less  prolonged  examination  of  an  object. 

The  difference  between  the  dry,  water,  and  oil  immersion  systems 
may  be  illustrated,  as  Frankel  has  pointed  out,  by  a  very  simple 
experiment.  If  a  glass  rod  is  inserted  into  an  empty  test-tube,  it 
is  easily  visible  owing  to  the  difference  in  refraction  between  the 
glass  and  the  surrounding  air.  If  the  tube  is  filled  up  with 
water  the  rod  is  seen  with  difficulty,  and  if,  instead  of  water,  ceclar 
oil  is  used,  the  part  of  the  rod  immersed  in  the  oil  will  entirely 
•disappear  from  view.  The .  rays  of  light  pass  through  an  optically 
uniform  medium  in  the  experiment  with  cedar  oil,  and  no  refraction 
or  reflection  of  rays  of  light  can  occur. 

To  use  an  oil  immersion  objective,  a  minute  drop  of  cedar  oil 
is  placed  on  the  centre  of  the  cover-glass,  and  the  lens  lowered 
by  means  of  the  coarse  adjustment  until  it  touches  the  oil.  The 
specimen  is  then  carefully  brought  into  focus  with  the  fine  adjust- 
ment. If  the  slide  is  held  between  the  finger  and  thumb  of  one 
hand,  and  moved  from  side  to  side  while  the  other  hand  is  working 
the  fine  adjustment,  there  can  be  no  danger  of  injuring  either  the 
objective  or  the  specimen. 

Microscopes  are  made  upon  either  the  Ross  or  the  Jackson 
model.  In  the  Ross  model  the  body  of  the  microscope  is  fixed 
at  its  base  to  a  transverse  arm,  which  is  raised  or  lowered  with 
it  by  the  rack  and  pinion.  In  the  Jackson,  model  the  body  is 
supported  for  a  great  part  of  its  length  on  a  solid  "  limb." 

In  the  Ross  model,  unless  the  body  and  transverse  arm  are  very 
solid  as  in  Powell  and  Lealand's  microscopes  (Fig.  23),  there  will  be 
vibration  at  the  ocular  end ;  but  in  the  Jackson  model  vibration  is 
practically  prevented,  and  this  is  most  essential,  especially  in  working 
with  very  high  powers. 

The  steadiness  of  the  microscope  also  largely  depends  upon  the 
form  of  stand.  There  are  four  different  types  of  stands.  The 
tripod  (Fig.  23) ;  the  plate,  with  double  columns ;  the  single  column, 
ending  in  a  plate  or  a  bent  claw;  and  the  horse-shoe  (Fig.  18). 

The  tripod  stand  with  cork  feet  is  the  steadiest  form  of  stand, 
but  it  is  cumbrous  and  expensive,  and  these  objections  also  apply  to 
the  model  made  by  Ross. 

The  single  upright  should  be  unquestionably  condemned,  as  it 


THE  BACTERIOLOGICAL  MICROSCOPE. 


71 


FIG.  16.— ENGLISH  MODEL. 


72  BACTERIOLOGY. 

freely  admits  of  vibration,  and  is  most  inconvenient  for  laboratory 
work,  The  heavy  horse-shoe  form  is  compact  and  firm,  and  the 
weight  of  it  can  hardly  be  considered  an  objection. 

The  tubular  body  is  from  eight  to  ten  inches  in  length,  and  within 
it  is  a  draw-tube  with  engraved  scale.  By  extending  the  draw-tube 
greater  magnification  is  obtained;  but  as  this  is  at  the  cost  of 
definition  it  should  hardly  ever  be  used  in  the  examination  of 
bacteria. 

A  triple  nose-piece  is  a  great  convenience,  saving  the  time  which 
is  otherwise  spent  in  replacing  objectives  of  different  magnifying 
power,  and  there  is  less  risk  of  injuring  them. 

Focus  should  be  obtained  by  means  of  a  rack  and  pinion  coarse 


FIG.  17.  —  REMOVABLE  MECHANICAL  STAGE. 

adjustment.  The  sliding  tube  is  not  to  be  recommended,  as  the 
motion  may  be  stiff,  encouraging  the  use  of  force,  which  in  turn  may 
result  in  the  objective  being  brought  violently  into  contact  with  the 
specimen,  injuring  the  lens  or  damaging  the  preparation ;  or  it  may 
get  too  loose  and  readily  slip  out  of  focus. 

The  stage  should  be  flat  and  rigid,  either  rectangular  or  circular, 
so  long  as  it  is  sufficiently  large  to  accommodate  a  plate- cultivation. 
A  removable  mechanical  stage  is  of  great  advantage  for  working 
with  high  powers,  as  a  motile  bacterium  can  be  constantly  kept  in 
view  while  one  hand  is  engaged  in  working  the  fine  adjustment 
(Fig.  17).  It  may  also  be  employed  as  a  finder  if  it  is  engraved 
with  a  longitudinal  and  vertical  scale,  and  provided  with  a  stop. 
The  mechanical  stage  must  be  removable,  so  that  the  stage  proper 


THE  BACTERIOLOGICAL  MICROSCOPE. 


73 


FIG.  18.— CONTINENTAL  MODEL. 


74  BACTERIOLOGY. 

may  be  free  from  any  attachments  when  required  for  the  examina- 
tion of  cultures. 

.  Diaphragms  are  necessary  for  regulating  the  amount  of  light. 
The  plan  of  using  a  series  of  discs,  with  apertures  of  different  sizes, 
should  be  avoided,  as  they  are  easily  lost,  and  bacteriological  investi- 
gations may  have  to  be  made  under  conditions  in  which  it  is  difficult 
to  replace  them.  A  better  plan  is  a  revolving  plate  with  apertures 
of  different  sizes,  but  the  most  convenient  form  is  the  iris  diaphragm 
(Fig.  19). 

The  sub-stage  condenser  is  quite  as  necessary  in  bacteriological 
work  as  a  high-power  objective.  In  fact,  the  condenser  and  the 
objective  should  be  considered  as  forming  one  optical  apparatus,  and 

the  microscope  regarded  quite  as 
incomplete  without  a  condenser  as 
it  would  be  without  an  objective. 

By  means  of  the  sub- stage  con- 
denser (Fig.  20)  the  rays  of  light 
are  concentrated  at  one  point  or 
on  one  particular  bacterium  ;  and 
for  the  best  definition  it  is  essential 
that  there  should  be  mechanical 

F,o.  m-lBiB  D.APHRAGM.  arrangements  for  accurately  cen- 

tring  and  focussing  the  condenser. 
It  may  even  with  advantage  be  provided  with  a  fine  adjustment. 

To  sum  up,  a  microscope  for  bacteriological  investigation  should 
be  provided  with  (1)  a  steady  stand  of  either  the  tripod  or  horse- 
shoe form ;  (2)  a  tubular  body  on  the  Jackson  model ;  (3)  a  wide- 
angled  sub-stage  condenser,  such  as  Abbe's;  (4)  objectives  of  an  inch, 
£th  of  an  inch,  and  a  T\th  homogeneous  immersion ;  (5)  a  removable 
mechanical  stage  ;  and  for  the  most  accurate  work  there  should  be 
centring  arrangements  and  a  coarse  and  fine  adjustment  to  an  oil- 
immersion  sub-stage  condenser  such  as  Powell  and  Lea-land's,  and 
a  i\th  homogeneous  oil-immersion  apo-chromatic  objective. 

With  regard  to  the  choice  of  a  microscope,  it  is  chiefly  a 
question  of  price.  The  most  perfect  instrument  is  the  large  model 
by  Powell  and  Lealand,  but  it  is  most  expensive,  and  quite  unsuit- 
able for  laboratory  work.  For  general  use  excellent  instruments 
are  made  by  Zeiss,  Leitz,  Reichert,  or  Swift.  The  bacteriological 
microscopes  of  these  makers  are  in  the  necessary  equipment 
practically  identical.  The  Zeiss  microscope  is  the  most  finished,  and 
costs  about  twenty  pounds.  A  similar  microscope  by  Leitz  and 
by  Swift  costs  about  eighteen,  and  both  make  an  excellent  students' 


THE  BACTERIOLOGICAL  MICROSCOPE. 


75 


bacteriological  microscope,  with  a  cheap  form  of  adjustment  to  the 
sub-stage  condenser,  at  a  total  cost  of  about  fifteen  pounds. 

Method  of  Illumination.— Good  daylight  is  the  best  for 
general  work.  The  microscope  should  be  placed  near  a  window 
with  a  northern  aspect.  Direct  sunlight  should  never  be  utilised,  and 
the  best  light  is  that  reflected  from  a  white  cloud.  When  daylight 
is  not  available  good  results  can  be  obtained  with  either  gas  or  a 


FIG.  20.— ABBE'S  CONDENSER  CONSTRUCTED  BY  ZEISS. 

paraffin  lamp.  In  the  author's  laboratory  the  microscope  lamps 
are  fitted  with  Welsbach  incandescent  mantles.  These  have  many 
advantages  over  an  Argand  burner  or  a  paraffin  lamp.  A  steady 
and  beautifully  white  light  is  obtained,  and  the  lamps  are  quickly 
lit,  and  require  comparatively  little  attention.  In  using  high  powers 
and  carefully  focussing  the  sub-stage  condenser,  the  image  of  the 
fabric  of  the  mantle  is  embarrassing,  and  is  an  objection  to  this 
light  for  the  most  accurate  observations,  but  in  other  respects,  and 


76 


BACTERIOLOGY. 


for  general  use,  it  is  the  best  form  of  artificial  illumination  for  the 
microscope. 

An  ordinary  paraffin  lamp  of  the  cheapest  form  may  be  used, 
but  there  are  many  objections  to  it,  such  as  the  shape  of  the 
chimney,  and  the  strise  and  defects  in  the  glass.  The  best  form  of 
paraffin  lamp  is  constructed  by  Baker  and  by  Swift  from  sug- 


FIG.  21. — MICROSCOPE  LAMP. 

gestions  by  Nelson  and  Dallinger  (Fig.  21) ;  there  is  also  a  similar 
but  much  larger  pattern  which  is  made  by  Swift  (Fig.  22).  This 
form  of  lamp  has  a  large  flat  bowl  for  the  oil.  It  is  attached  to 
a  standard,  and  can  be  raised  or  lowered  to  the  desired  position. 
The  chimney  is  of  metal  and  blackened,  so  that  there  is  no  reflected 
light,  and  it  may  also  with  advantage  be  provided  with  a  shade, 
so  that  no  light  reaches  the  eye  except  through  the  microscope. 

The  burner  may  be  made  to  revolve,  so  that  either  the  edge  or 


THE    BACTERIOLOGICAL   MICROSCOPE.  77 

the  flat  of  the  flame  may  be  utilised.  Great  care  should  be  taken 
to  have  the  wick  evenly  trimmed.  The  best  paraffin  oil  should 
be  burnt,  and  it  is  as  well  to  add  a  small  lump  of  camphor.  The 
metal  chimney  has  an  aperture  in  front,  giving  exit  to  the  rays  of 
light,  which  is  closed  in  by  a  slip  of  glass.  The  glass  is  very  liable 
to  crack  when  exposed  to  the  full  force  of  the  flame,  and  it  is  as  well, 
therefore,  to  be  provided  with  a  stock  of  glass  slips,  which  have 


FIG.  22. — LARGE  MICROSCOPE  LAMP. 

been  annealed  by  being  enveloped  in  a  cloth  and  boiled  for  two  or 
three  hours. 

The  flat  of  the  flame  is  used  with  low  powers.  The  image  of 
the  flame  is  reflected  by  a  plane  mirror,  and  a  bull's-eye  condenser 
interposed  between  the  lamp  and  the  mirror  to  give  an  equal 
illumination  of  the  whole  field.  In  working  with  high  powers  the 
lamp  is  turned  with  the  flame  edgewise,  and  the  mirror  is  dispensed 
with.  By  working,  as  it  is  termed,  directly  on  the  edge  of  the 
flame,  the  illumination  is  greatly  increased,  and  a  band  of  light  can 


78  BACTERIOLOGY. 

be  concentrated  on  that  part  of  the  microscopical  preparation  which 
requires  most  careful  study  (Fig.  23). 

To  obtain  the  best  definition  considerable  time  must  be  spent  in 
the  arrangement  of  the  illumination.  The  lamp  and  microscope 
having  been  placed  in  position,  a  low  power  is  first  used  and  the 
smallest  diaphragm.  On  looking  through  the  microscope  it  will 
probably  be  observed  that  the  image  of  the  diaphragm  is  not  in 
the  centre  of  the  field.  By  moving  the  centring  screw  of  the  con- 
denser this  may  be  adjusted.  The  image  of  the  edge  of  the  flame 
may  not  be  central,  and  this  must  be  adjusted  by  moving  the  lamp 
into  position.  The  low  power  is  then  replaced  by  a  high  power, 
the  largest  diaphragm  used,  and  the  bacteria  brought  into  focus. 
The  diaphragm  must  now  be  replaced  by  one  of  medium  size,  and 
by  racking  the  condenser  up  and  down,  a  point  will  be  arrived  at 
when  the  image  of  the  edge  of  the  flame  appears  as  an  intensely 
bright  band  of  light.  If  this  is  not  exactly  in  the  centre  of  the 
field  the  centring  screws  of  the  condenser  must  again  be  adjusted. 
Lastly,  by  trying  different  sizes  of  diaphragms,  and  focussing  with 
the  fine  adjustment,  and  using  the  correction  collar,  we  arrive  at  the 
sharpest  possible  image  of  the  bacteria. 

When  the  condenser  has  been  accurately  centred,  it  will  still  be 
necessary  to  focus  it  for  each  individual  specimen,  so  as  to  correct 
for  difference  in  the  thickness  of  slides  and  the  layers  of  mounting 
medium.  Correction  for  different  thickness  of  cover-glasses  must 
in  each  case  be  made  by  means  of  the  collar  adjustment  in  the  follow- 
ing way.  A  high-power  eye-piece  is  substituted  for  the  ordinary 
eye-piece,  and  the  fault  in  the  image  will  thereby  be  intensified.  By 
moving  the  collar  completely  round,  first  in  one  direction  and  then 
the  other,  while  carefully  observing  the  effect  on  the  image,  it  will 
be  seen  to  become  obviously  worse  whichever  way  the  collar  is  turned. 
The  collar  must  then  be  turned  through  gradually  diminishing  dis- 
tances until  an  intermediate  point  is  reached  at  which  the  best  image 
results  with  the  high-power  eye-piece,  and  on  replacing  this  by  the 
low-power  eye-piece  the  sharpest  possible  image  will  be  obtained. 

Effect  of  the  sub-stage  condenser. — The  sub-stage  condenser 
gives  the  most  powerful  illumination  when  it  has  been  racked  up 
until  it  almost  touches  the  specimen.  It  produces  a  cone  of  rays  of 
very  short  focus,  and  the  apex  of  the  cone  should  correspond  with  the 
particular  bacterium  or  group  of  bacteria  under  observation.  The 
effect  of  the  condenser  without  a  diaphragm  is  to  obliterate  what 
Koch  has  termed  the  structure  picture.  If  the  component  parts  of  a 
tissue  section  were  colourless  and  of  the  same  refractive  power  as 


THE    BACTERIOLOGICAL   MICROSCOPE. 


79 


80  BACTERIOLOGY. 

the  medium  in  which  the  section  is  mounted,  nothing  would  be 
visible  under  the  microscope.  As,  however,  the  cells  and  their  nuclei, 
and  the  tissue  fibres  do  differ  in  this  respect,  the  rays  which  pass 
through  them  are  diffracted,  and  an  image  of  lines  and  shadows  is 
developed.  If  in  such  a  tissue  there  were  minute  coloured  objects, 
and  if  it  were  possible  to  mount  the  tissue  in  a  medium  of  exactly 
the  same  refractive  power,  the  tissue  being  then  invisible,  the 
detection  of  the  coloured  objects  would  be  much  more  easy.  This 
is  exactly  what  is  required  in  dealing  with  bacteria  which  have  been 
stained  with  aniline  dyes,  and  the  desired  result  can  be  obtained 
by  the  use  of  the  sub-stage  condenser. 

If  we  use  the  full  aperture  of  the  condenser  the  greatly  converged 
rays  play  on  the  component  parts  of  the  tissue,  light  enters  from 


FIG.  24.— RAMSDEX  MICROMETER  EYE-PIECE. 

all  sides,  the  shadows  disappear,  and  the  structure  picture  is  lost. 
If  now  a  diaphragm  is  inserted,  so  that  we  are  practically  only 
dealing  with  parallel  rays,  the  structure  picture  reappears.  As  the 
diaphragm  is  gradually  increased  in  size  the  structure  picture 
gradually  becomes  less  and  less  distinct,  while  the  colour  picture, 
the  image  of  the  stained  bacteria,  becomes  more  and  more  intense. 
When,  therefore,  bacteria  in  the  living  condition  and  unstained  tissues 
are  examined  a  diaphragm  must  be  used,  and  when  attention  is 
to  be  concentrated  upon  the  stained  bacteria  in  a  section  or  in  a 
cover-glass  preparation,  the  diaphragm  must  be  removed  and  the 
field  flooded  with  light. 

Micrometer. — For  the  measurement  of  bacteria  a  stage  micro- 
meter may  be  used  with  a  camera  lucida.  The  stage  micrometer 
consists  of  a  slip  of  thin  glass  ruled  with  a  scale  consisting  of  tenths 
and  hundredths  of  a  millimetre.  The  image  of  this  can  be  projected 


THE  BACTERIOLOGICAL  MICROSCOPE. 


81 


on  a  piece  of  paper,  and  a  drawing  made,  and  the  object  to  be 
measured  can  then  be  projected  on  the  paper  and  compared  with  the 
scale. 

In  the  Ramsden  micrometer  eye-piece  (Fig.  24)  two  fine  wires 
are  stretched  across  the  field  of  an  eye -piece,  one  of  which  can  be 
moved  by  a  micrometer  screw.  In  the  field  there  is  also  a  scale 
with  teeth,  and  the  interval  between  them  corresponds  to  that  of  the 
threads  of  the  screw.  The  circumference  of  the  brass  head  is  usually 
divided  into  one  hundred  parts,  and  a  screw  with  one  hundred  threads 
to  the  inch  is  used.  The  bacterium  to  be  measured  is  brought  into  a 


FIG.  25.— MICROMETER  EYE-PIECE  BY  ZEISS. 

position  in  which  one  edge  appears  to  be  in  contact  with  the  fixed 
wire,  and  the  micrometer  screw  is  turned  until  the  travelling  wire 
appears  to  be  in  contact  with  the  other  edge.  The  scale  in  the 
field  and  the  scale  on  the  milled  head  together  give  the  number  of 
complete  turns  of  the  screw  and  the  value  of  a  fraction  of  a  turn  in 
separating  the  wires. 

In  the  micrometer  eye-piece  constructed  by  Zeiss,  the  eye- piece 
with  a  glass  plate  with  crossed  lines  is  carried  across  the  field  by 
means  of  a  micrometer  screw  (Fig.  25).  Each  division  on  the  edge 
of  the  drum  corresponds  to  *01  mm.  Complete  revolutions  of  the 
drum  are  counted  by  means  of  a  figured  scale  in  the  visual  field. 
Another  method  of  measuring  bacteria  will  be  referred  to  in  the 

6 


82  BACTERIOLOGY. 

chapter  on  micro -photography.  The  unit  of  measurement  is  one 
thousandth  of  a  millimetre  or  a  macro-millimetre  or  micron,  and  is 
expressed  by  the  Greek  letter  p.. 

CARE    OF   THE    MICROSCOPE. 

After  use  the  objectives,  sub-stage  condenser,  and  eye-piece 
should  be  carefully  wiped  with  soft  linen,  an  old  silk  handkerchief, 
or  chamois  leather,  and  the  microscope  covered  with  a  bell-glass  to 
protect  it  from  dust.  If  a  lens  comes  into  contact  with  Canada 
balsam  it  must  be  very  carefully  wiped  with  a  soft  rag  moistened 
with  alcohol,  and  then  cleaned  with  a  soft  leather.  Microscopes 
should  not  be  exposed  to  the  fumes  of  sulphuretted  hydrogen, 
chlorine,  or  volatile  acids. 


CHAPTER    VIII. 

MICROSCOPICAL   EXAMINATION    OF   BACTERIA. 

(A)  BACTERIA  IN  LIQUIDS,  CULTURES,  AND  FRESH  TISSUES. 

IN  conducting  bacteriological  researches  the  importance  of  absolute 
cleanliness  cannot  be  too  strongly  insisted  upon.  All  instru- 
ments, glass  vessels,  slides,  and  cover-glasses  should  be  thoroughly 
cleansed  before  use.  A  wide-mouthed  glass  jar  should  always  be 
close  at  hand,  containing  refuse  alcohol  for  the  reception  of  re- 
jected slide  preparations  or  dirty  cover-glasses.  When  required 
again  for  use,  slides  can  be  easily  wiped  clean  with  a  soft  rag.  Cover- 
glasses  require  further  treatment,  for,  unless  they  are  perfectly 
clean,  it  is  difficult  to  avoid  the  presence  of  air  bubbles  when 
mounting  specimens.  They  should  be  left  in  strong  acid  (hydro- 
chloric, sulphuric,  or  nitric)  for  some  hours  ;  they  are  then  washed, 
first  with  water  and  then  with  alcohol,  and  carefully  wiped  with  a 
soft  rag.  The  same  principle  applies  in  the  preparation  and 
employment  of  culture  media ;  any  laxity  in  the  processes  of 
sterilisation,  or  insufficient  attention  to  minute  technical  details, 
will  surely  be  followed  with  disappointing  results  by  contamination 
of  the  cultures,  resulting  in  the  loss  of  much  time. 

For  the  preparation  of  microscopical  specimens  it  will  be  found 
convenient  to  use  a  platinum  inoculating  needle.  This  consists  of 
two  or  three  inches  of  platinum  wire  fused  into  the  end  of  a  glass 
rod  about  eight  inches  in  length.  Platinum  is  employed  as  it 
rapidly  cools  after  being  raised  to  a  white  heat  in  the  flame  of  a 
Bunsen  burner.  It  is  thus  completely  sterilised,  and  in  a  few 
moments  is  cool  enough  not  to  destroy  the  bacteria  with  which  it  is 
brought  into  contact. 

When  using  platinum  needles,  either  for  inoculating  fresh  tubes 
in  carrying  on  a  series  of  pure  cultures,  or  in  transferring  a  small 
portion  of  a  cultivation  to  a  cover- glass  for  examination  under  the 
microscope,  the  careful  sterilisation  of  the  needle  by  heating  the 

83 


84  BACTERIOLOGY. 

platinum  wire  till  it  is  white  hot  in  every  part,  and  heating  also 
as  much  of  the  glass  rod  as  is  made  to  enter  the  test-tube,  must 
be  carried  out  with  scrupulous  care.  Indeed  it  is  a  good  plan  to 


FIG.  26. — INOCULATING  NEEDLES. 

let  it  become  a/orce  of  habit  to  sterilise  the  needle  before  and  after 
use  on  every  occasion,  whatever  may  be  the  purposes  for  which  it 
is  employed. 

UNSTAINED  BACTERIA. 

The  bacteria  in  liquids,  such  as  pus,  blood,  and  culture- fluids,  can 
be  investigated  in  the  unstained  condition  by  transferring  a  drop  with 
a,  looped  platinum  needle  or  a  capillary  pipette  to  a  slide,  covering 
it  with  a  clean  cover-glass,  and  examining  without  further  treat- 
ment. If  it  is  desirable  to  keep  the  specimen  under  prolonged 
observation,  a  drop  of  sterilised  water  or  salt  solution  must  be  run  in 
at  the  margin  of  the  cover-glass  to  counteract  the  tendency  to  dry. 

Cultures  on  solid  media  can  be  examined  by  transferring  a  small 
portion  with  a  sterilised  needle  to  a  drop  of  sterilised  water  on  a 
slide,  thinning  it  out,  and  covering  with  a  cover-glass  as  already 
described. 

Tissues  in  the  fresh  state  may  be  teased  out  with  needles  in 
sterilised  salt  solution,  and  pressed  out  into  a  sufficiently  thin  layer 
between  the  slide  and  cover-glass.  Glycerine  may  in  many  cases 
be  substituted  for  salt  solution,  especially  for  the  examination  of 
micro-organisms  such  as  Actinomyces  and  mould  fungi. 

There  is,  as  a  rule,  no  difficulty  in  recognising  the  larger  micro- 
organisms such  as  those  just  mentioned;  but  when  we  have  to 
deal  with  very  small  bacilli  and  micrococci,  they  may  possibly  be 
mistaken  for  granular  detritus  or  fat-crystals,  or  vice  versa.  They 
are  distinguished  by  the  fact  that  fatty  and  albuminous  granules 
are  altered  or  dispersed  by  acetic  acid,  and  changed  by  solution 
of  potash ;  alcohol,  chloroform,  and  ether  dissolve  out  fat-crystals 


MICROSCOPICAL   EXAMINATION   OF   BACTERIA.  85 

or  fatty  particles ;  on  the  other  hand,  micro-organisms  remain 
unaffected  by  these  reagents.  Baurngarten  demonstrated  tubercle 
bacilli  in  sections  by  treating  them  with  potash,  which  clarified 
the  tissues  and  brought  the  bacilli  clearly  into  view.  Actinomyces 
and  other  vegetable  structures  will  not  disappear  when  sections  are 
immersed  in  weak  hydrochloric  acid  and  mounted  in  glycerine. 

In  examining  unstained  bacteria,  it  is  necessary,  in  order  to 
obtain  the  structure  picture,  that  the  light  entering  the  microscope 
should  be  reduced  by  employing  a  small  diaphragm,  and  the  sub-stage 
condenser  carefully  centred  and  focussed.  To  focus  an  unstained 
specimen  in  which  only  bacteria  are  present,  is  often  difficult.  The 
slide  may  be  gently  raised  towards  the  objective,  and  the  stage 
may  be  constructed  to  enable  this  to  be  done  with  the  index  finger 
(Fig.  1 6).  If  on  tilting  the  slide  the  organisms  come  into  focus  it 
will  serve  as  a  guide  in  working  the  fine  adjustment.  Another  plan 
when  bacteria  are  examined  in  water,  is  to  look  for  an  air-bubble, 
and  then  to  focus  its  edge  until  the  bacteria  appear  in  view. 

The  simple  method  of  covering  the  liquid  with  a  cover-glass  will 
not  answer  for  a  prolonged  examination,  as  the  liquid  evaporates  and 
the  specimen  dries  up.  To  keep  living  bacteria  under  observation 
for  any  length  of  time,  in  order  to  study  their  movements  or  spore- 
formation,  a  special  slide  must  be  employed  (p.  120). 

STAINED  BACTERIA. 

Weigert  first  pointed  out  the  value  of  the  aniline  dyes  for 
staining  bacteria,  and  we  are  principally  indebted  to  Koch,  Ehrlich, 
Gram,  and  Lo flier  for  many  valuable  processes. 

The  staining  of  fresh  preparations,  especially  those  with  no 
coagulable  albumen  to  fix  them,  may  be  carried  out  by  the  method 
of  His.  A  slide  is  prepared  as  already  described  for  the  exami- 
nation of  micro-organisms  in  the  fresh  state.  The  reagents  are 
then  applied  by  placing  them  with  a  pipette  drop  by  drop  at 
one  margin  of  the  cover-glass,  and  causing  them  to  flow  through 
the  preparation  by  means  of  a  strip  of  filter-paper  placed  at  the 
opposite  margin. 

Babes  recommends  another  rapid  means  of  examining  cultivations. 
A  little  of  the  growth,  removed  by  means  of  a  sterilised  platinum 
hook  or  small  loop,  is  spread  out  on  a  cover-glass  into  as  thin  a 
film  as  possible  :  when  almost  dry,  a  drop  or  two  of  a  weak  aqueous 
solution  of  methyl  violet  is  allowed  to  fall  from  a  pipette  upon  the 
tilm.  The  cover-glass  with  the  drop  of  stain  is,  after  a  minute, 


86  BACTERIOLOGY. 

carefully  turned  over  on  to  a  slide,  and  the  excess  of  stain  gently 
and  gradually  removed  by  pressure  with  a  strip  of  filter-paper. 
This  affords  a  rapid  means  of  demonstration — for  example,  of  a 
cultivation  of  Koch's  comma  bacilli  in  nutrient  gelatine — enabling 
the  microbes  to  be  seen  in  some  parts  of  the  preparation  both 
stained  and  in  active  movement. 

COVER-GLASS  PREPARATIONS. 

Bacteria  may  be  spread  out  into  a  thin  layer  on  a  cover-glass, 
and  then  treated  with  a  dye,  or  sections  of  tissues  containing  bacteria 
can  be  stained  and  then  mounted  in  the  usual  way. 

The  method  of  making  a  cover-glass  preparation  is  one  which  is 
very  commonly  employed.  In  addition  to  its  value  as  a  means  of 
examining  bacteria  in  liquids  and  solid  culture  media,  it  affords 
the  additional  advantage  of  enabling,  if  necessary,  a  large  number 
of  preparations  to  be  made,  which,  when  dried,  can  be  preserved, 
stained  or  unstained,  in  ordinary  cover-glass  boxes ;  they  are 
then  in  a  convenient  form  for  transport,  and  can  be  mounted 
permanently  at  leisure. 

The  method  is  as  follows  :  A  cover-glass  is  smeared  with  the 
cut  surface  of  an  organ  or  pathological  growth,  or  with  sputum  ; 
or  a  drop  of  blood,  pus,  or  culture- fluid  is  conveyed  to  it  with  a 
looped  platinum  needle.  It  is  absolutely  necessary  to  spread  out 
the  micro-organisms  into  a  sufficiently  thin  layer,  so  that  the 
individual  bacteria  may  be  as  much  as  possible  in  the  same  plane, 
otherwise  some  in  the  field  will  be  in  focus  and  others  out  of 
focus,  and  it  would  be  impossible  to  obtain  a  satisfactory  photograph 
of  such  a  specimen.  To  overcome  this  it  will  be  necessary,  in  the 
case  of  cultures  on  solid  media,  to  diffuse  the  bacteria  in  a  little 
sterilised  water ;  and  even  cultures  in  liquids  may  sometimes  with 
advantage  be  diluted  in  the  same  way.  By  means  of  another 
cover-glass  the  juice  or  fluid  is  squeezed  out  between  them  into  a 
thin  layer,  and  on  sliding  them  apart  each  cover-glass  bears  on  one 
side  a  thin  film  of  the  material  to  be  examined  ;  or  a  culture  is 
spread  out  into  a  thin  film  by  means  of  a  hooked  platinum  needle. 
The  cover- glass  is  then  placed  with  the  prepared  side  upwards,  and 
allowed  to  dry.  After  a  few  minutes,  it  is  taken  up  with  a  pair  of 
flat-bladed  or  spring  forceps,  with  the  prepared  side  uppermost,  and 
passed  rapidly  from  above  downwards  three  times  through  the 
flame  of  a  spirit  lamp  or  Bunsen  burner.  Two  or  three  drops  of 
an  aqueous  solution  of  fuchsine  or  methyl  violet  will  be  sufficient  to 
cover  the  film,  and  after  a  minute  or  two  the  surplus  stain  is  washed 


MICROSCOPICAL   EXAMINATION   OF   BACTERIA.  87 

off  with  distilled  water  by  means  of  a  siphon  apparatus  or  a  wash- 
bottle.  The  cover-glass  may  be  allowed  to  dry,  and  then  mounted 
in  Canada  balsam,  or  it  may,  while  still  wet,  be  turned  over  on  to  a 
slide,  the  excess  of  water  removed  with  filter-paper,  and  the  exposed 
surface  wiped  dry.  It  may  first  be  examined  with  a  power  of  about 
250  diams. ;  and  if  a  high  magnification  is  required,  which  is  usually 
the  case,  a  droplet  of  cedar  oil  is  placed  on  the  cover-glass,  and  the 
specimen  examined  with  an  immersion  lens. 

If  the  specimen  is  to  be  made  permanent,  fix  the  cover-glass  at 
one  corner  with  the  thumb,  and  with  a  soft  rag  carefully  wipe  off 
the  cedar  oil ;  then  float  off  the  cover-glass  by  running  in  distilled 
water  at  its  margin,  and  having  made  a  little  ledge  with  a  strip  of 
filter- paper,  place  the  cover- glass  up  against  it  upon  one  of  its 
edges  and  leave  it  to  dry.  When  perfectly  dry  mount  in  Canada 
balsam,  or  put  it  away  in  a  cover-glass  box  provided  with  a  label  of 
contents. 

In  many  cases  it  is  necessary  or  preferable  to  apply  the  stain 
for  a  much  longer  period.  This  may  best  be  effected  by  pouring 
some  of  the  staining  solution  into  a  watch-glass,  and  allowing  the 
cover- glasses  to  swim  on  the  surface,  with  their  prepared  side,  of 
course,  downwards.  Throughout  all  these  manipulations  it  is 
necessary  to  bear  in  mind  which  is  the  prepared  surface  of  the 
cover-glass. 

Instead  of  using  the  watery  solutions  of  the  aniline  dyes  the 
author  prefers  in  many  cases  to  use  stronger  solutions,  and  to  reduce 
the  staining  by  a  momentary  immersion  in  alcohol.  Very  beautiful 
preparations  of  streptococci,  sarcinse  and  other  bacteria  can  be 
obtained  by  this  method,  which  is  as  follows  :  Cover-glass  prepara- 
tions are  stained  with  carbolised  fuchsine  (Neelsen's  solution)  for 
about  two  minutes,  rinsed  in  alcohol  for  a  few  seconds,  quickly 
washed  in  water,  and  either  examined  in  water  or  dried  and 
mounted  in  the  usual  way.  The  extent  of  decolorisation  is  a 
matter  of  practice  :  a  momentary  immersion  in  alcohol  is  sometimes 
sufficient ;  too  long  immersion  will  remove  too  much  of  the  colour ; 
too  short  immersion  will  leave  the  delicate  outlines  indistinct.  This 
method  is  especially  valuable  for  sarcinae  and  streptococci,  the 
divisions  between  the  elements  being  sharply  defined,  and  as  any 
albuminous  particles  or  debris  in  the  preparation  are  decolorised, 
much  cleaner  and  sharper  preparations  are  obtained  than  with  the 
watery  solutions.  Loffler's  and  other  concentrated  solutions  may 
also  be  used,  but  Neelsen's  solution  may  be  regarded  as  the  standard 
one  for  this  method. 


88  BACTERIOLOGY. 

Aniline  oil,  carbolic  acid,  and  some  other  chemicals,  when  added 
to  the  aniline  dyes,  have  the  property  of  acting  in  the  manner 
of  mordants,  in  some  way  fixing  the  colour  in  the  bacteria,  so  that 
they  are  not  so  readily  acted  upon  by  decolorising  agents. 

Lojfler's  /Solution. — Potash  intensifies  the  staining  power,  and 
Koch  and  Lb'ffler  have  both  used  it  with  methylene  blue.  Loffler's 
solution  consists  of  30  grammes  of  methylene  blue  in  100  grammes 
of  1  in  10,000  solution  of  potash.  It  may  be  used  with  advantage 
for  almost  all  kinds  of  bacteria. 

Gram's  Method.— With  a  solution  of  gentian-violet  the  whole 
film  on  the  cover-glass  is  at  first  stained  violet.  By  immersing  the 
cover-glass  in  a  solution  of  iodine  in  iodide  of  potassium  the  stain 
is  fixed  in  the  bacilli,  but  not  in  any  debris,  pus  cells,  or  tissue 
elements  present  in  the  film.  Consequently  by  transferring  the- 
cover-glass  to  alcohol  the  bacilli  alone  remain  stained,  the  violet 
colour  being  merely  changed  to  blue.  By  employing  a  contrast 
colour,  such  as  eosin,  a  double  staining  is  obtained.  In  some 
bacteria  the  sheath  is  by  this  method  differentiated  from  the 
protoplasmic  contents. 

The  stock  solution  of  gentian-violet  is  prepared  by  shaking  up 
1  cc.  of  pure  aniline  with  twenty  parts  of  distilled  water,  and 
filtering  the  emulsion.  Half  a  gramme  of  the  best  finely  powdered 
gentian- violet  is  dissolved  in  the  clear  filtrate,  and  the  solution  filtered 
before  use. 

The  details  of  the  method  will  now  be  described.  In  the  first 
place,  it  is  much  better  to  employ  the  aniline-gentian-violet  solution 
quite  freshly  prepared,  and  the  following  useful  method  is  invariably 
used  by  the  author :  Place  four  or  five  drops  of  pure  aniline  in 
a  test-tube,  fill  it  three-quarters  full  with  distilled  water,  close  the 
mouth  of  the  tube  with  the  thumb,  and  shake  it  up  thoroughly. 
Filter  the  emulsion  twice,  and  pour  the  filtrate  into  a  watch-glass 
or  glass  capsule.  To  the  perfectly  clear  aniline -water  thus  obtained 
add  drop  by  drop  a  concentrated  alcoholic  solution  of  gentian -violet 
till  precipitation  commences.  Cover-glasses  must  be  left  in  this 
solution  about  ten  minutes,  transferred  to  iodiiie-potassic-iodide 
solution  until  in  itwo  or  three  minutes  the  film  becomes  uniformly 
brown,  and  then  rinsed  in  alcohol.  The  process  of  decolorisation  may 
be  hastened  by  dipping  the  cover-glass  in  clove-oil  and  returning  it 
again  to  alcohol.  The  cover-glass  is  once  more  immersed  in  clove-oilr 
then  dried  by  gently  pressing  between  two  layers  of  filter-paper, 
and  finally  mounted  in  Canada  balsam. 


MICROSCOPICAL  EXAMINATION  OF  BACTERIA.  89 

DOUBLE  STAINING  OF  COVER-GLASS  PREPARATIONS. 

To  double  stain  cover-glass  preparations  they  can  be  treated  by 
Ehrlich's  method  for  staining  tubercular  sputum,  or  by  Neelsen's 
modification,  or  by  staining  with  eosin  after  treatment  by  the 
method  of  Gram. 

Ehrlich's  method  is  as  follows :  Five  parts  of  aniline  oil  are 
shaken  up  with  one  hundred  parts  of  distilled  water,  and  the 
emulsion  filtered  through  moistened  filter-paper.  A  saturated 
alcoholic  solution  of  fuchsine,  methyl-violet,  or  gentian -violet,  is 
added  to  the  filtrate  in  a  watch-glass,  drop  by  drop,  until  precipitation 
commences.  Weigert  recommended  that  exactly  eleven  parts  of  the 
dye  should  be  used  to  one  hundred  parts  of  the  aniline  solution. 

Cover-glass  preparations  are  floated  in  this  mixture  for  fifteen 
minutes  to  half  an  hour,  then  washed  for  a  few  seconds  in  dilute 
nitric  acid  (one  part  nitric  acid  to  two  of  water),  and  then  rinsed 
in  distilled  water.  The  stain  is  removed  from  everything  except 
the  bacilli;  but  the  ground  substance  can  be  after-stained  brown 
if  the  bacilli  are  violet,  or  blue  if  they  have  been  stained  red. 

yeelsen's  Solution  and  Methylene  Blue. — Ziehl  suggested  the  use  of 
carbolic  acid  as  a  substitute  for  aniline  oil,  and  Neelsen  recommended 
a  solution  composed  of  100  cc.  of  a  5  per  cent,  watery  solution 
of  carbolic  acid,  10  cc.  of  absolute  alcohol,  and  1  gramme  of 
fuchsine.  This  stain  is  commonly  known  as  the  Neelsen  or  Ziehl - 
Neelseii  solution.  Cover-glass  preparations  are  floated  on  the  hot 
dye  for  two  minutes,  they  are  then  rinsed  in  dilute  sulphuric  acid 
25  per  cent.,  washed  in  water,  immersed  in  watery  solution  of 
methylene  blue  for  three  minutes,  again  washed  in  water,  dried, 
and  mounted  in  balsam. 

Gram's  Solution  and  Eosin. — Double  staining  of  cover-glasses  can 
be  obtained  by  combining  Gram's  method  with  eosin.  The  method 
is  very  useful  for  differentiating  the  sheath  of  Streptococcus 
pyogenes  and  Bacillus  anthracis,  from  the  protoplasmic  contents, 
and  for  staining  preparations  of  pneumonic  sputum,  or  of  micrococci 
and  other  micro-organisms  in  pus.  After  decolorising  the  prepara- 
tion in  alcohol,  the  cover-glass  is  transferred  to  a  weak  solution 
of  eosin  for  two  or  three  minutes,  then  washed  again  in  alcohol, 
immersed  in  clove-oil,  dried  between  filter-paper,  and  mounted  in 
balsam. 

STAINING  OF  SPORES. 

A  slight  modification  of  the  ordinary  process  employed  in  making 
cover-glass  preparations  has  to  be  adopted  to  stain  the  spores  of 


90  BACTERIOLOGY. 

bacilli.  Under  ordinary  circumstances  the  stain  will  not  penetrate 
the  sheath,  but  if  it  can  be  made  to  penetrate,  it  is  not  readily 
removed.  The  cover-glass  preparation  must  be  heated  to  a  tem- 
perature of  210°  C.,  for  half  an  hour,  or  passed  as  many  as  twelve 
times  through  the  flame  of  a  Bunsen  burner,  or  exposed  to  the 
action  of  strong  sulphuric  acid  for  several  seconds,  and  then  a  few 
drops  of  a  watery  solution,  of  an  aniline  dye  may  be  applied  in  the 
usual  way. 

To  double  stain  spore- bearing  bacilli  the  cover-glass  preparations 
may  be  floated,  for  from  twenty  minutes  to  an  hour,  on  Ehrlich's 
fuchsine-aniline-water,  or  on  the  Ziehl-Neelsen  solution.  The  stain 
must  be  heated — by  preference  in  a  capsule  placed  in  a  sand-bath — 
until  steam  rises.  The  fuchsine  is  removed  from  the  bacilli  by 
rinsing  in  water  and  washing  in  weak  hydrochloric  acid,  and  then 
the  preparations  are  washed  again  in  water,  and  floated  for  a  few 
minutes  on  a  watery  solution  of  methylene  blue.  They  are  again 
rinsed  in  water,  dried,  and  mounted.  Neisser's  decolorising  solution 
consists  of  25  parts  of  hydrochloric  acid  to  75  parts  of  alcohol. 

STAINING  OF  FLAGELLA. 

Koch  first  stained  flagella  by  floating  the  cover-glasses  on  a 
watery  solution  of  hsematoxylin.  From  this  they  were  transferred 
to  a  5  per  cent,  solution  of  chromic  acid,  or  to  Miiller's  fluid,  by 
which  the  flagella  obtain  a  brownish- black  coloration.  The  author 
succeeded  in  demonstrating  and  photographing  flagella  in  prepara- 
tions stained  with  a  saturated  solution  of  gentian  violet  in  absolute 
alcohol ;  but  these  methods  are  now  superseded  owing  to  the  much 
more  satisfactory  method  introduced  by  Loffler. 

Loffler's  method  depends  upon  the  employment  of  a  mordant. 
Loffler  tried  tannate  of  iron,  and  after  a  number  of  experiments 
the  following  method  was  introduced.  An  aqueous  solution  of 
ferrous  sulphate  is  added  to  an  aqueous  solution  of  tannin  (20  per 
cent.),  until  the  mixture  turns  a  violet-black  colour,  then  3  or  4  cc. 
of  a  1  in  8  aqueous  solution  of  logwood  are  added.  This  constitutes 
the  mordant,  and  a  few  drops  of  carbolic  acid  may  be  added,  and  the 
solution  kept  in  well-stoppered  bottles.  The  dye  consists  of  1  cc.  of 
a  1  per  cent,  solution  of  caustic-soda,  added  to  100  cc.  of  aniline 
water,  in  which  4  or  5  grammes  of  either  methyl  violet,  methylene  blue, 
or  fuchsine,  are  dissolved.  A  cover-glass  preparation  is  made  in 
the  ordinary  way,  the  bacteria  being  diffused  in  water,  and  then 
spread  out  in  a  very  thin  film.  After  drying  and  very  carefully 
fixing,  the  film  is  covered  with  the  mordant,  and  the  cover-glass 


MICROSCOPICAL  EXAMDFATTOV  Of  BACTERIA.  91 

held  over  the  flame  until  steam  rises.  The  mordant  is  then  washed 
off  with  distilled  water,  and  all  traces  removed  from  the  edge  of 
the  cover-glass  with  alcohoL  The  stain  is  filtered,  and  a  few  drops 
allowed  to  fall  on  the  film,  and  after  a  few  minutes  the  cover- 
very  carefully  warmed  until  steam  rises.  The  stain 
off  with  distilled  water,  and  is  ready  to  be  examined  and 
subsequently  mounted.  For  some  bacteria  it  is  necessary  to  modify 
the  solutions,  either  by  the  addition  of  acetic  or  sulphuric  acid,  or 
by  varying  the  quantity  of  soda  solution. 

L  introduced  a  modification  of  Lofflers  system.  Cover- 
are  floated  for  from  two  to  twelve  hours  on  a  solution 
of  1  per  cent,  tannin  and  |  per  cent,  hydrochloric  acid. 
After  washing  in  water  the  preparation  is  stained  with  a  saturated 
alcoholic  solution  of  any  of  the  amtine  dyes  diluted  in  the  propor- 
tion of  2  drops  of  the  dye  to  20  of  water.  The  cover-glasses 
in  the  solution  for  from  two  to  four  hours  are  then 
in  water,  and  examined.  The  best  results  are  obtained  with 
carbofised  fnchsine,  diluted  in  the  proportion  of  2  drops  to  20  drops 
of  1  per  cent,  carbolic.  Trenkmann  also  recommended  the  use  of 
catechu  and  logwood  as  mordants,  with  the  addition  of  very  dilute 
acid,  and  subsequent  «taimt»g  with  fnchsine. 

Lutesch  suggested  the  use  of  ferric  acetate.  To  avoid  any 
deposit  on  the  surface  of  the  preparation,  freshly  prepared  saturated 
ferric  acetate  is  used,  and  5  to  10  drops  of  acetic  add  are  added  to 
16  ce.  of  the  mordant.  After  warming  the  solution  the  preparation 
is  washed  in  water,  followed  by  20  per  cent,  acetic  acid, 


gentian-violet  in  aniling*  water. 

Tan  Ermengem  used  a  mordant  composed  of  1  part  of 
2  per  cent,  solution  of  osmie  acid,  2  parts  of  10  to  25  per  cent, 
solution  of  tannin,  with  to  cvtsj^  100  ce.  of  this  mixture  4  or  5  drops 
of  acetic  acid.  A  black  ink  is  thus  formed,  and  the  solution  is 
applied  for  from  five  to  thirty  minutes.  After  washing  in  water  and 
alcohol  the  cover-glasses  are  placed  in  a  solution  of  nitrate  of  silver 
and  transferred  to  another  solution  composed  of  5  grammes  of  gallic 
acid,  3  grammes  of  tannin,  10  grammes  of  acetate  of  soda,  and  330 
grammes  of  distilled  water.  In  a  few  moments  they  are  again  placed 
in  nitrate  of  silver,  and  then  washed  and  mounted  in  balsam. 

oCuLVO  i>  tPf^MlOd  AOSWCsTS   ^ndl  lOB*  CCB^tstMTl    HI  if sffEmffP|W ^IflffBttfv        AO<6? 

preparations  are  left  for  one  minute  in  solution  of  tannin,  washed  in 
dirtilipd  water,  transferred  for  a  minute  to  50  per  cent,  phospho- 
molybdic  acid,  again  washed  and  stained  from  three  to  five 


92  BACTERIOLOGY. 

in  hot  saturated  solution  of  f  uchsine  in  aniline  water,  washed  in  watery 
dried  on  filter  paper,  and  mounted  in  balsam.  The  tannin  solution 
consists  of  1  part  of  tannin  to  100  cc.  of  50  per  cent,  alcohol. 

Nicolle  and  Morax  also,  have  modified  Loffler's  method.  Per- 
fectly clean  cover-glasses  are  used,  and  the  film  is  dried  without 
fixing  in  the  flame.  Cover-glasses  are  covered  with  the  mordant, 
and  heated  for  about  ten  seconds,  and  when  steam  rises  the  mordant 
is  shaken  off  and  the  film  rinsed  with  water.  The  same  process  is 
repeated  three  or  four  times,  and  finally  the  cover-glass  is  stained 
with  Neelsen's  solution,  holding  it  over  the  flame  once  or  twice 
for  a  quarter  of  a  minute  ;  it  is  then  washed  and  examined. 

Bunge  prefers  as  a  mordant  a  mixture  of  aqueous  solution  of 
tannin  with  1  in  20  aqueous  solution  of  sesquichloride  of  iron  in  the 
proportion  of  3  parts  of  the  tannin  solution,  1  part  of  the  iron 
solution,  with  the  addition  of  1  cc.  of  a  saturated  watery  solution  of 
f  uchsine  added  to  10  cc.  of  the  mixture.  The  mordant  is  kept  before 
use,  and  applied  for  five  minutes.  The  preparation  is  then  washed  and 
stained  with  ISTeelsen's  solution.  In  another  plan  the  cover-glasses 
are  immersed  for  one  half  to  one  minute  in  5  per  cent,  solution  of 
acetic  acid,  washed  and  dried.  The  mordant  is  then  applied  three  or 
four  times,  and  the  cover-glasses  washed,  dried,  and  then  stained  with 
gentian-violet,  dipped  in  1  per  cent,  acetic  acid,  washed,  dried,  and 
mounted.  Peroxide  of  hydrogen  may  be  added  to  the  mordant r 
drop  by  drop ;  it  becomes  reddish-brown  in  colour,  and  must  be 
shaken  up  and  filtered  before  use.  Cover-glasses  are  exposed  to  its 
action  for  about  a  minute,  and  Neelsen's  solution  is  used  for 
staining. 

Hessert  dispenses  with  the  mordant.  The  film  is  fixed  by 
treating  cover-glasses  with  a  saturated  alcoholic  solution  of  corrosive 
sublimate.  .After  washing,  the  cover-glass  is  stained  for  thirty  to 
forty  minutes  in  a  hot  dye,  by  preference  a  10  per  cent,  watery 
solution  of  saturated  alcoholic  solution  of  fuchsine. 

COVER-GLASS  IMPRESSIONS. 

One  of  the  most  instructive  methods  for  examining  micro- 
organisms is  to  make  an  impression-penetration.  This  enables  us, 
in  many  cases,  to  study  the  relative  position  of  individual  micro- 
organisms one  to  another  in  their  growth  on  solid  cultivating  media, 
and  in  some  cases  produces  the  most  exquisite  preparations  for  the 
microscope.  A  perfectly  clean,  usually  small-sized,  cover-glass  is 
carefully  deposited  on  a  plate-cultivation,  and  gently  and  evenly 
pressed  down.  One  edge  is  then  carefully  levered  up,  with  a  needle, 


MICROSCOPICAL   EXAMINATION    OF   BACTERIA.  93 

and  the  cover-glass  lifted  off  by  means  of  forceps.  It  is  then 
allowed  to  dry,  passed  through  the  flame  three  times,  and  stained 
as  already  described.  In  some  cases  of  plate- cultures,  especially 
where  no  liquefaction  has  taken  place,  the  growth  is  bodily  trans- 
ferred to  the  cover-glass,  and  a  vacant  area  left  on  the  gelatine 
or  agar-agar,  corresponding  exactly  with  the  form  and  size  of  the 
cover-glass  employed. 

PRESERVATION  OF  PREPARATIONS. 

After  examining  a  cover-glass  preparation  with  an  oil  immersion 
objective  the  cedar  oil  must  be  carefully  wiped  off,  and  the  slide 
set  aside  for  the  Canada  balsam  to  set.  At  a  convenient  time  all 
preparations  should  be  sealed  with  a  ring  of  Hollis'  glue ;  the 
cedar  oil  used  at  subsequent  examinations  of  the  specimen  will 
not  be  able  to  work  its  way  under  the  cover-glass,  and  prevent 
the  balsam  from  hardening.  When  it  is  ringed  cedar  oil  can  be 
readily  wiped  off,  and  the  specimen  cleaned  without  danger  of 
moving  the  cover-glass  and  injuring  the  preparation. 

(B)  BACTERIA  IN  SECTIONS  OF  TISSUES. 

Methods  of  Hardening  and  Decalcifying  Tissues. — To  harden  small 
organs,  such  as  the  viscera  of  a  mouse,  they  should  be  placed  on 
a  piece  of  filter- paper  at  the  bottom  of  a  small  wide -mouthed  glass 
jar,  and  covered  with  about  twenty  times  their  volume  of  absolute 
alcohol.  Larger  organs,  pathological  growths,  etc.,  are  treated  in 
the  same  way,  but  must  first  be  cut  into  small  pieces,  or  cubes, 
varying  from  a  quarter  of  an  inch  to  an  inch  in  size.  Miiller's 
fluid  may  also  be  employed,  and  methylated  spirit  may  be  sub- 
stituted for  alcohol,  from  motives  of  economy.  Tissues  hardened  in 
absolute  alcohol  are  ready  for  cutting  in  two  or  three  days,  and 
those  hardened  in  Miiller's  fluid  in  as  many  weeks. 

Teeth,  or  osseous  structures,  must  first  be  placed  in  a  decalcifying 
solution,  such  as  Kleinenberg's.  When  sufficiently  softened,  they 
are  allowed  to  soak  in  water,  to  wash  out  the  picric  acid,  and  then 
transferred  through  weak  spirit  to  absolute  alcohol.  Ebner's  solu- 
tion also  gives  excellent  results,  especially  when  the  structures  to 
be  decalcified  are  placed  in  fresh  solution  from  time  to  time. 

Methods  of  Embedding,  Fixing,  and  Cutting. — The  author 
finds  that  freezing  with  ether  combined  with  the  method  of  em- 
bedding in  celloidin  gives  excellent  results.  The  pieces  of  tissue 
to  be  embedded  are  placed,  after  the  process  of  hardening  is  com- 


94  BACTERIOLOGY, 

pleted,  in  a  mixture  of  ether  and  alcohol  for  an  hour  or  more. 
They  are  then  transferred  to  a  solution  of  celloiclin  in  equal  parts 
of  ether  and  alcohol,  and  left  there,  usually  for  several  hours. 

The  piece  of  tissue  is  then  placed  in  a  glass  capsule,  and  some 
of  the  celloidin  solution  poured  over  it.     The  capsule  can  be  placed 


FIG.  27. — SWIFT'S  FREEZING  MICROTOME. 

bodily  in  60  to  80  per  cent,  alcohol,  and  left  until  the  following 
morning.  The  celloidin  will  then  be  of  the  consistency  of  wax. 
The  piece  of  tissue  is  next  cut  out,  and  after  trimming  off  superfluous 
celloidin  is  put  in  water  until  it  sinks.  It  is  then  transferred  to 
gum,  and  frozen  and  cut  with  a  freezing  microtome. 

For  cutting  with  Jung's  microtome,  the  tissues  are  embedded 


MICROSCOPICAL   EXAMINATION   OF   BACTERIA. 


95 


in  paraffine  or  celloidin,  and  mounted  on  cork ;  or,  if  firm  enough, 
they  may  be  fixed  upon  cork  without  any  embedding  material  at 
all.  Paraffine,  dissolved  in  chloroform,  will  be  found  very  service- 
able as  an  embedding  material. 

Corks  ready  cut  for  the  clamp  of  the  microtome  are  smeared 
over  with  the  solution  of  celloidin.  This  can  be  applied  with  a 
glass  rod  to  the  surface  which  is  to  receive  the  piece  of  tissue- 
The  corks  are  then  set  aside  for  the  film  of  celloidin  to  harden. 
In  the  case  of  lung,  or  degenerated  broken-down  tissue,  the 
specimen  should  be  left  for  a  much  longer  time  than  is  found  to 
be  sufficient  for  firmer  structures.  When  ready,  it  is  removed 
from  the  celloidin  solution  with  forceps  and  placed  upon  the  pre- 


FIG.  28. — JUNG'S  MICROTOME. 

pared  cork.  Enough  of  the  solution,  which  is  of  syrupy  consistence, 
is  allowed  to  fall  on  the  piece  of  tissue  to  cover  it  completely,  and 
the  mounted  specimen  is  placed  in  the  alcohol  to  harden.  The 
specimen  will  be  ready  for  cutting  next  day. 

The  specimen  may  be  more  neatly  embedded  by  fixing  it  with 
a  pin  in  a  small  paper  tray,  pouring  the  celloidin  solution  over  it, 
and  then  placing  the  tray  in  alcohol  to  harden  the  celloidin.  The 
embedded  specimen  is  then  fixed  on  a  cork,  which  has  been  cut  for 
the  clamp  of  the  microtome.  The  celloidin  in  the  section  disappears 
in  the  process  of  clearing  with  clove-oil. 

In  the  case  of  specimens  embedded  in  celloidin,  or  mounted 
directly  on  a  cork,  the  tissue,  as  well  as  the  blade  of  the  knife,  should 
be  kept  constantly  bathed  with  alcohol,  and  the  sections  transferred 
from  the  blade  with  a  camel's-hair  brush,  and  floated  in  alcohol. 


96  BACTERIOLOGY. 

For  fixing  directly  on  cork,  small  organs  and  pieces  of  firm  tissue 
such  as  the  kidneys  of  a  mouse,  or  liver,  we  may  employ  gelatine  or 
glycerine  gelatine,  liquefied  over  a  Bunsen  burner  in  a  porcelain 
capsule.  Glycerine  gelatine  may  be  used  with  advantage  for  fixing 
irregular  pieces  of  tissue,  as  it  does  not  become  of  a  consistency 
that  would  injure  the  edge  of  the  knife.  The  cork,  with  specimen 
affixed,  is  placed  in  alcohol,  and  is  ready  for  cutting  sections  next 
day. 

Material  infiltrated  with  paraffine  must  be  cut  perfectly  dry, 
and  the  sections  prevented  from  rolling  up  by  gentle  manipulation 
with  a  camel's-hair  brush.  They  must  then  be  picked  off  the  blade 
of  the  knife  with  a  clean  needle,  and  dropped  into  a  watch-glass 
containing  xylol.  This  dissolves  out  the  paraffine.  The  sections  are 
then  transferred  to  alcohol  to  get  rid  of  the  xylol,  and  then  to  the 
staining  solution. 

Staining  Bacteria  in  Tissue  Sections, — Sections  of  fresh  tissues 
made  with  the  freezing  microtome  are  to  be  floated  in  '8  per  cent, 
salt  solution,  and  then  carefully  transferred,  well  spread  out  on  a 
platinum  lifter,  to  a  watch-glass  containing  absolute  alcohol.  Simi- 
larly, sections  selected  from  those  cut  with  Jung's  microtome  may 
be  transferred  from  the  spirit  to  absolute  alcohol.  The  sections 
may  be  then  stained  by  any  of  the  methods  to  be  described. 

It  is  often  advisable  to  employ  some  method  which  will  enable 
one  to  study  the  structure  of  the  tissue  itself ;  and  sections,  however 
stained,  should  always  be  first  examined  with  low  powers,  to  enable 
one  to  recognise  the  tissue  under  examination,  and  to  examine  in 
many  cases  the  topographical  distribution  of  masses  of  bacteria. 
With  a  power  of  about  250  diams.  (one-sixth),  very  many  bacteria 
can  be  distinguished ;  and  with  the  oil  immersion  lenses  the  minutest 
bacilli  and  micrococci  can  be  recognised,  and  the  exact  form  of 
individual  bacteria  accurately  determined.  As  most  good  modern 
instruments  are  provided  with  a  triple  nose-piece,  there  is  no  loss 
of  time  in  examining  a  preparation  successively  with  these  different 
powers. 

Weigert's  Method. — A  very  useful  method  for  staining  both 
the  tissue  and  the  bacteria  is  as  follows :  Place  the  sections  for 
from  six  to  eighteen  hours  in  a  1  per  cent,  watery  solution  of  any  of 
the  basic  aniline  dyes  (methyl  violet,  gentian  violet,  fuchsine,  Bis- 
marck brown).  To  hasten  the  process,  place  the  capsule  containing 
the  solution  in  the  incubator,  or  heat  it  to  45°  C.  A  stronger 
solution  may  also  be  employed,  in  which  case  the  sections  are  far 
more  rapidly  stained,  and  are  easily  over-stained.  In  the  latter  case 


MICROSCOPICAL   EXAMINATION   OF   BACTERIA.  97 

they  must  be  treated  with  a  half-saturated  solution  of  carbonate  of 
potash.  In  either  case  the  sections  are  next  washed  with  distilled 
water,  and  passed  through  60  per  cent,  alcohol  into  absolute  alcohol. 
When  almost  decolorised,  spread  out  the  section  carefully  on  a 
platinum  lifter  and  transfer  it  to  clove-oil,  or  stain  with  picro-carmine 
solution  (Weigert's)  for  half  an  hour,  wash  in  water,  alcohol,  and 
then  treat  with  clove-oil.  After  the  final  treatment  with  clove- oil, 
transfer  with  the  platinum  lifter  to  a  clean  glass  slide.  Dry  the 
preparation  by  pressure  with  a  piece  of  filter-paper  folded  several 
times,  and  preserve  in  Canada  balsam,  dissolved  in  xylol. 

Gram's  Method. — In  the  method  of  Gram  sections  are  stained 
for  ten  minutes  in  a  capsule  containing  aniline-gentian- violet  solution. 
Great  care  must  be  taken  not  to  injure  the  sections.  If  there  is 
any  difficulty  in  finding  them,  it  is  best  to  carefully  pour  off  the 
stain  and  fill  up  the  capsule  with  water.  The  sections  are  then  readily 
visible,  and  can  be  taken  up  on  the  end  of  a  glass  rod  and  placed 
in  the  iodine  and  iodide  of  potassium  solution,  where  they  remain  for 
two  or  three  minutes,  until  stained  uniformly  brown  and  resembling 
in  appearance  a  boiled  tea-leaf.  They  are  then  placed  in  absolute 
alcohol,  and  washed  by  carefully  moving  the  sections  in  the  liquid 
with  a  glass  rod.  When  completely  decolorised  they  are  spread  out 
on  a  lifter,  and  transferred  to  clove-oil  until  completely  clarified. 
Each  is  transferred  with  a  lifter  to  a  slide,  and  the  clove-oil  is 
run  off  and  then  completely  removed  by  gently  pressing  two  or 
three  layers  of  filter-paper  upon  the  section.  Finally,  the  section 
is  mounted  in  Canada  balsam. 

The  process  of  decolorisation  may  be  hastened  by  transferring  the 
section  from  alcohol  to  clove-oil,  and  back  again  to  alcohol,  repeating 
this  two  or  three  times. 

On  examination  the  tissue  appears  colourless,  or  slightly  tinged 
yellow  from  too  long  immersion  in  the  iodine  solution,  while  the 
micro-organisms  are  stained  blue  or  blue-black. 

Double  staining  is  obtained  by  transferring  the  sections  after 
decolorisation  to  eosin,  Bismarck  brown,  or  vesuvin.  They  are  left 
in  a  watery  solution  for  two  or  three  minutes,  then  again  washed  in 
alcohol,  before  clarifying  in  clove-oil  and  mounting  in  balsam. 

Another  instructive  method  is  to  place  the  decolorised  sections 
in  picro-carminate  of  ammonia  for  three  or  four  minutes,  and  then 
treat  with  alcohol  and  clove-oil. 

A  similar  result  is  obtained  by  placing  the  sections  in  Orth's 
solution  (picro-lithium  carmine),  transferring  to  acidulated  alcohol, 
and  then  passing  through  clove-oil  and  mounting  in  balsam. 

7 


98  BACTERIOLOGY. 

In  Ehrlich's  method  delicate  sections  are  liable  to  be  injured  by 
immersion  in  the  nitric  acid,  and  therefore  Watson- Oheyne  suggested 
the  use  of  formic  acid. ' 

The  Ziehl-Neelsen  method,  in  which  sulphuric  acid  is  used  instead 
of  nitric  acid,  is  much  to  be  preferred  to  Ehrlich's  method. 

Ziehl-Neelsen  Method. — The  solution  is  warmed,  and  sections 
left  in  it  for  ten  minutes.  The  red  colour,  which  disappears  when 
the  section  is  placed  in  weak  sulphuric  acid  (25  per  cent.),  may 
partly  return  when  the  section  is  placed  in  water.  In  this  case  the 
section  must  be  again  immersed  in  acid  and  passed  backwards  and 
forwards  from  acid  to  water  until  the  red  colour  has  completely,  or 
almost  completely,  disappeared.  It  must  be  thoroughly  washed  in 
water  to  remove  all  traces  of  the  acid,  and  then  placed  in  a  watery 
solution  of  methylene  blue  for  two  or  three  minutes,  washed  again 
in  water,  immersed  in  alcohol,  clarified  in  clove-oil,  and  mounted  in 
the  usual  way.  Sections  are  brilliantly  stained,  and  the  results  are 
very  permanent. 

Many  special  methods  of  staining  have  been  introduced,  and  will 
be  given  in  subsequent  chapters  with  the  description  of  the  bacteria 
to  which  they  apply.  The  methods  already  described  are  those 
which  are  more  or  less  in  constant  use  in  studying  bacteria  and  in 
conducting  original  researches. 


CHAPTER   IX. 

PREPARATION   OF   NUTRIENT    MEDIA    AND    METHODS    OF 
CULTIVATION. 

To  cultivate  micro-organisms  artificially,  and,  in  the  case  of  the 
pathogenic  bacteria,  to  fulfil  the  second  of  Koch's  postulates,  they 
must  be  supplied  with  nutrient  material  free  from  pre-existing 
micro-organisms'.  Hitherto  various  kinds  of  nutrient  liquids  have 
been  employed,  and  in  many  cases  they  still  continue  to  be 
used  with  advantage,  but  for  general  use  they  have  been,  in  a 
great  measure,  supplanted  by  the  methods  of  cultivation  on  sterile 
solid  media  about  to  be  described.  The  advantages  of  the  latter 
methods  are  numerous.  In  the  first  place,  in  the  case  of  liquid 
media,  in  spite  of  elaborate  precautions  and  the  expenditure  of  much 
labour  and  time,  it  was  almost  impossible  or  extremely  difficult  to 
obtain  a  pure  culture.  When  a  drop  of  liquid  containing  several  kinds 
of  bacteria  is  introduced  into  a  liquid  medium,  we  have  a  mixed 
cultivation  from  the  very  first.  If  in  the  struggle  for  existence 
some  bacteria  were  unable  to  develop  in  the  presence  of  others,  or 
a  change  of  temperature  and  soil  allowed  one  form  to  predominate 
over  another,  then  we  might  be  led  to  the  conclusion  that  many 
bacteria  were  but  developmental  forms  of  one  and  the  same  micro- 
organism ;  while  possibly  the  contamination  of  such  cultures  might 
lead  to  the  belief  in  the  transformation  of  a  harmless  into  a  patho- 
genic bacterium.  The  secret  of  the  success  of  Koch's  methods  greatly 
depends  upon  the  possibility,  in  the  case  of  starting  with  a  mixture 
of  micro-organisms,  of  being  able  to  isolate  them  completely  one 
from  another,  and  to  obtain  an  absolutely  pure  growth  of  each 
cultivable  species.  When  sterile  nutrient  gelatine  has  been  liquefied 
in  a  tube  and  inoculated  with  a  mixture  of  bacteria  in  such  a  way 
that  the  individual  micro-organisms  are  distributed  throughout  it, 
and  the  liquid  is  poured  out  on  a  plate  of  glass  and  allowed  to  solidify, 
the  individual  bacteria,  instead  of  moving  about  freely  as  in  a  liquid 
medium,  are  fixed  in  one  spot,  where  they  develop  individuals  of 

99 


DESCRIPTION    OF    PLATE    II. 
Pure-cultivations  of  Bacteria. 

FIG.  1. — In  the  depth  of  Nutrient  Gelatine.  A  pure-cultivation  of  Kochs 
comma-bacillus  (Spirillum  choleras  Asiatics)  showing  in  the  track  of 
the  needle  a  funnel-shaped  area  of  liquefaction  enclosing  an  air-bubble, 
and  a  white  thread.  Similar  appearances  are  produced  in  cultivations  of 
the  comma-bacillus  of  Metchnikoff. 

FIG.  2. —  On  the  surface  of  Nutrient  Gelatine.  A  pure-cultivation  of  Bacillus 
typhosus  on  the  surface  of  obliquely  solidified  nutrient  gelatine. 

FIG.  3. —  On  the  surface  of  Nutrient  Agar-agar.  •  Pure-cultivation  of  Bacillus 
indicus  on  the  surface  of  obliquely  solidified  nutrient  agar-agar.  The 
growth  has  the  colour  of  red  sealing-wax,  and  a  peculiar  crinkled 
appearance.  After  some  days  it  loses  its  bright  colour  and  becomes 
purplish,  like  an  old  cultivation  of  Micrococcus  prodigiosus. 

FIG.  4. — On  the  surface  of  Nutrient  Agar-agar.  A  pure-cultivation  obtained 
from  an  abscess  (Staphylococcus  pyogenes  aureus). 

FIG.  5. —  On  tJie  surface  of  Nutrient  Agar-agar.  A  pure-cultivation  obtained 
from  green  pus  (Bacillus  pyocyaneus).  The  growth  forms  a  whitish, 
transparent  layer,  composed  of  slender  bacilli,  and  the  green  pigment 
is  diffused  throughout  the  nutrient  jelly.  The  growth  appears  green  by 
transmitted  light,  owing  to  the  colour  of  the  jelly  behind  it. 

FIG.  (5. —  On  the  surface  of  Potato.  A  pure-cultivation  of  the  bacillus  of 
glanders  on  the  surface  of  sterilised  potato. 


Plate  TL 


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OF  THB 

TJNIVERSITY 


NUTRIENT   MEDIA    AND   METHODS   OF    CULTIVATION. 


101 


squeezing  through  a  linen  cloth  or  a  meat  press.  The  red  juice  thus 
obtained  must  be  brought  up  to  a  litre  by  transferring  it  to  a  large 
measuring  glass  and  adding  distilled  water.  It  is  then  poured 
into  a  sufficiently  large  and  strong  beaker,  and  set  aside  after  the 
addition  of  10  grammes  of  peptone,  5  grammes  of  common  salt  and 
100  grammes  of  best  gelatine. 

In  about  half  an  hour  the  gelatine  is  sufficiently  softened,  and 
subsequent    heating   in    a    water-bath    causes  it   to   be   completely 


FIG.  30.— HOT  Am  STERILISKK. 


dissolved.  The  danger  of  breaking  the  beaker  may  be  avoided  by 
placing  a  cloth,  several  times  folded,  at  the  bottom  of  the  water-bath. 

The  next  process  requires  the  greatest  care  and  attention.  Some 
micro-organisms  grow  best  in  a  slightly  acid,  others  in  a  neutral 
or  slightly  alkaline,  medium.  For  example,  for  the  growth  and 
characteristic  appearances  of  the  comma  bacillus  of  Asiatic  cholera 
a  faintly  alkaline  soil  is  absolutely  essential.  This  slightly  alkaline 
medium  will  be  found  to  answer  best  for  most  micro-organisms,  and 
may  be  obtained  as  follows  :  — 

With  a  clean  glass  rod  dipped  in  the  mixture,  the  reaction 
upon  litmus- paper  may  be  ascertained,  and  a  concentrated  solution 
of  carbonate  of  soda  must  be  added  drop  by  drop,  until  red  litmus- 


102 


BACTERIOLOGY. 


paper  becomes  faintly  blue.     If  it  has  been  made  too  alkaline,  it  can 
be  neutralised  by  the  addition  of  lactic  acid. 

Finally,  the  mixture  is  heated  for  an  hour  in  the  water-bath. 
Ten  minutes  before  the  boiling  is  completed,  the  white  of  an  egg 
beaten  up  with  the  shell  is  added,  and  the  liquid  is  then  filtered 
while  hot.  For  the  filtration,  the  hot- water  apparatus  (Fig.  31) 
can  be  used  with  advantage,  furnished  with  a  filter  of  Swedish 

paper,    which  may  be  conveniently 
made  in  the  following  way  : — 

About  eighteen  inches  square 
of  the  best  and  stoutest  filter  paper 
is  first  folded  in  the  middle,  'and 
then  creased  into  sixteen  folds.  The 
filter  is  made  to  fit  the  glass  funnel 
by  gathering  up  the  folds  like  a  fan, 
and  cutting  off  the  superfluous  part. 
The  creasing  of  each  fold  should  be 
made  firmly  to  within  half  an  inch 
of  the  apex  of  the  filter,  which  part 
is  to  be  gently  inserted  into  the 
tube  of  the  funnel.  To  avoid 
bursting  the  filter  at  the  point,  the 
broth,  when  poured  out  from  the 
flask,  should  be  diiected  against 
the  side  of  the  filter  with  a  glass 
rod.  During  filtration  the  funnel 
should  be  covered  over  writh  a 
circular  plate  of  glass,  and  the  pro- 
cess of  filtration  must  be  repeated, 
if  necessary,  until  a  pale,  straw- 
coloured,  perfectly  transparent 
filtrate  results.- 

The  sterilised  test-tubes  are  filled  to  about  a  third  of  their  depth 
by  pouring  in  the  gelatine  carefully  and  steadily,  or  by  employing  a 
small  sterilised  glass  funnel.  The  object  of  this  care  is  to  prevent 
the  mixture  touching  the  part  of  the  tube  with  which  the  plug 
comes  into  contact ;  otherwise,  when  the  gelatine  sets,  the  cotton- 
wool adheres  to  the  tube  and  becomes  a  source  of  embarrassment  in 
subsequent  procedures.  As  the  tubes  are  filled  they  are  placed  in 
the  test-tube  basket,  and  must  then  be  sterilised.  They  are  either 
lowered  into  the  steam  steriliser,  when  the  thermometer  indicates 
100°  0.,  for  twelve  minutes  for  four  or  five  successive  days,  or  they 


FIG.  31. — HOT-WATER  FILTERING 
APPARATUS. 


NUTRIENT   MEDIA    AND   METHODS   OF    CULTIVATION. 


103 


may  be  transferred  to  the  test-tube  water-bath,  and  heated  for  an 
hour  a  day  for  three  successive  days. 

If  the  gelatine  shows  any  turbidity  after  these  processes  it  must 


FIG.  32.— METHOD  OF  MAKING  A  FOLDED  FILTER. 

be  poured  back  from  the  test-tubes  into  a  flask,  boiled  up  for  ten 
minutes,  and  filtered  once  more,  and  the  processes  of  sterilisation  just 
described  must  be  repeated. 


FIG.  33. — STEAM  STERILISED 


Nutrient  Agar-agar. — Agar-agar  is  a  substance  prepared 
from  seaweed  which  grows  on  the  coasts  of  Japan  and  India, 
and  is  supplied  in  long  crinkled  strips.  It  boils  at  90°  C.,  and 


104 


BACTERIOLOGY. 


remains  solid  up  to  a  temperature  of  about  45°  C.  It  is  there- 
fore substituted  for  gelatine  in  the  preparation  of  a  jelly  for  the 
cultivation  of  those  bacteria  which  will  only  grow,  or  grow  best,  in 
the  incubator  at  the  temperature  of  the  blood.  It  may  also  be 
employed  at  ordinary  temperatures  for  bacteria  which  liquefy 
gelatine.  The  preparation  is  conducted  on  much  the  same  principles 
as  those  already  described.  Instead,  however,  of  100  grammes  of 
gelatine,  only  about  20  grammes  of  agar-agar  are  employed  (1'5  to 
2  per  cent.),  and  to  facilitate  its  solution  it  must  be  allowed  to  soak 
in  salt  water  overnight.  For  the  filtration,  flannel  is  substituted 

for  filter- paper,  or  may 
be  used  in  combination 
with  the  latter.  The 
hot-water  apparatus 
is  invariably  employed, 
unless,  to  accelerate 
the  process,  the  glass 
funnel  and  receiver  are 
bodily  transferred  to 
the  steam  steriliser.  If 
the  conical  cap  cannot 
be  replaced,  cloths  laid 
over  the  mouth  of  the 
steriliser  must  be  em- 
ployed instead.  It  may 
be  necessary  to  repeat 
the  process  of  filtra- 
tion, but  it  must  not  be 
expected  that  such  a 
brilliant  transparency 
can  be  obtained  as  with 
gelatine.  The  final 

result,  when  solid,  should  be  colourless  and  clear;  but  if  slightly 
milky,  it  may  still  be  employed. 

A  little  liquid  gradually  collects  in  the  tubes,  being  expressed  by 
the  contraction  of  the  agar-agar. 

Wort-gelatine  is  used  in  studying  the  bacteria  of  fermentation. 
It  is  made  by  adding  from  5  to  10  per  cent,  of  gelatine  to  beer- wort. 
Glycerine  Agar-agar. — This  is  prepared  by  adding  5  per 
cent,  of  glycerine  to  nutrient  agar-agar,  after  the  boiling  and  before 
the  filtration,  and  other  modifications  can  be  made  for  special 
purposes  by  the  addition  of  grape-sugar  or  of  gelatine. 


FIG.  34. — INCUHATOR. 


NUTRIENT   MEDIA    AND   METHODS   OF   CULTIVATION.  105 

After  the  final  treatment  in  the  steam  steriliser  some  of  the 
of  gelatine  and  agar-agar  are  placed  upright  and  allowed  to 
set,  and  others  are  placed  on  an  inclined  plane  or  in  the  blood-serum 
in.Npissator,  and  left  to  gelatinise  with  an  oblique  surface. 

(B)  METHODS  OF  EMPLOYING  NUTRIENT  JELLY  IN  TEST-TUBES 
AND  ON  GLASS  PLATES. 

Test-tube-cultivations. — To    inoculate    test-tubes   containing 
nutrient  jelly,   the  cotton- wool  plug  is  first  twisted  round  in  case 
there  are  any  adhesions  between  the  plug  and  the  test-tube.     It  is 
then    removed    with    the    thumb    and 
index    finger   of  the   right    hand,    and 
placed    between    the  fourth   and  fifth 
fingers   of   the    left    hand,    instead    of 
being  put  down  on  the  laboratory  table 
and    thereby    probably    contaminated 
with  bacteria   or  the  spores  of  mould 
fungi.     A    sterilised    needle    charged, 
for    example,    with  blood  or  pus  con- 
taining bacteria,  or  with  a  colony  from 
a  plate-culture,  is  thrust  once  in  the 
middle    line   into   the   nutrient    jelly, 
and    steadily    withdrawn.       The    tube 
should    be    held   horizontally    or   with   • 
its  mouth  downward,  to  avoid,  as  far 
as   possible,    accidental   contamination 

from  the  gravitation  of  germs  in  the 

j  , ,  .  i  i       FIG.  35. — METHOD  OF  IXOCULAT- 

air;  and  the  plug  replaced  as  quickly       ^  A  TJMJT.TCBB   COXTAINING 

as  possible.      The  cotton- wool  project-        STERILE  NUTRIENT  JELLY. 
ing  beyond  the  mouth  of  the  tube  is 

then  thoroughly  burnt  in  the  flame  of  a  Bunsen  burner  or  blow- 
pipe, and  an  india-rubber  cap  fitted  over  the  mouth  of  the  tube 

The  chances  of  error  arising  from  contamination  of  the  culti- 
vations are  reduced  by  avoiding  draughts  at  the  time  of  inoculation, 
and  it  is  best  that  these  manipulations  should  be  carried  on  in  a 
quiet  room  in  which  the  tables  and  floor  are  wiped  with  damp  cloths, 
rather  than  in  a  laboratory  in  which  the  air  becomes  charged  with 
germs  through  constant  sweeping  and  dusting,  and  the  entrance 
and  exit  of  classes  of  students.  In  conducting  any  investigation 
a  dozen  or  more  tubes  should  be  inoculated,  and  if  by  chance  an 
adventitious  germ,  in  spite  of  all  precautions,  gains  an  entrance, 


106  BACTERIOLOGY. 

the  contaminated  tube  can  be  rejected,  and  the  experiments  con- 
tinued with  the  remaining  pure  cultivations. 

When,  however,  one  tube  containing  a  liquid  medium  is  in- 
oculated from  another,  as  in  the  process  of  preparing  plate-cultures, 
or  when  a  culture  is  made  from  a  tube  in  which  the  growth  has 
liquefied  the  gelatine,  it  is  obvious  that  the  tubes  cannot  be  inverted 
or  held  horizontally,  and  they  must  then  be  held  and  inoculated  as 
in  Fig.  38.  To  inoculate  those  tubes  of  nutrient  media  which  have 
been  solidified  obliquely,  the  point  of  a  straight  sterilised  needle 
charged  with  the  material  to  be  cultivated  is  traced  over  the  surface 
of  the  jelly  from  below  upwards,  or  the  inoculated  material  may  be 
spread  out  with  a  hooked  or  looped  needle. 

Examination  of  Test-tube-cultivations. — The  appearances  pro- 
duced by  the  growths  in  test-tubes  can  be  in  most  cases  sufficiently 
examined  with  the  naked  eye.  In  some  cases  the  jelly  is  partially 
or  completely  liquefied,  while  in  others  it  remains  solid.  The 
growths  may  be  abundant  or  scanty,  coloured  or  colourless.  The 
nutrient  jelly  may  itself  be  tinged  or  stained  with  products  resulting 
from  the  growth  of  the  organisms.  When  liquefaction  slowly  takes 
place  in  the  nesdle  track,  or  the  organism  grows  without  producing 
this  change,  the  appearances  which  result  are  often  very  delicate, 
and  in  some  cases  very  characteristic.  The  appearance  of  a  simple 
white  thread,  of  a  central  thread  with  branching  lateral  filaments, 
of  a  cloudiness,  or  of  a  string  of  beads  in  the  track  of  the  needle, 
may  be  given  as  examples. 

In  some  cases  much  may  be  learnt  by  examining  the  growth  with 
a  magnifying  glass.  Here,  however,  a  difficulty  may  be  encountered, 
for  the  cylindrical  form  of  the  tube  so  distorts  the  appearance  of  its 
contents,  that  the  examination  is  rendered  somewhat  difficult.  To 
obviate  this,  a  very  simple  contrivance  may  be  employed  with 
advantage.  This  consists  of  a  rectangular  vessel,  about  four  inches 
in  height  and  two  inches  in  width,  which  may  be  easily  constructed 
by  cementing  together  two  slips  of  glass  to  form  the  back  and  front, 
with  three  slips  of  stout  glass  with  ground  edges  forming  the  sides 
and  base  (Cheshire).  The  front  may  be  constructed  of  thin  glass, 
and  the  base  of  the  vessel  made  to  slope  so  that  the  test-tube  when 
placed  in  the  vessel  has  a  tendency  to  be  near  the  front.  The 
vessel  is  filled  with  a  mixture  of  the  same  refractive  index  as  the 
nutrient  gelatine.  The  latter  has  a  refractive  index  rather  higher 
than  water,  which  is  about  1'333  ;  alcohol  has  a  ref reactive  index  of 
1'374.  The  vessel  is  filled  with  water,  and  alcohol  is  then  added 
until  the  proper  density  is  reached.  The  test-tube  is  placed  in  the 


NUTRIENT   MEDIA   AND   METHODS    OF   CULTIVATION. 


107 


vessel,  and  held  in  position  by  means  of  a  clip.  The  vessel  can  be 
fixed  on  the  inclined  stage  of  the  microscope,  and  the  contents  of  the 
tube  conveniently  examined  with  low-power  objectives. 

Plate- cultivations. — By  this  method,  as  already  mentioned,  a 
mixture  of  bacteria,  whether  in  fluids,  excreta,  or  in  cultivations  on 
solid  media,  can  be  so  treated  that  the  different  species  are  isolated 
one  from  the  other,  and  perfectly  pure  cultivations  of  each  of  the 
cultivable  bacteria  in  the  original  mixture  established  in  various 
nutrient  media.  We  are  enabled  also  to  examine  under  a  low 
power  of  the  microscope  the  individual  colonies  of  bacteria,  and  to 
distinguish  by  their  characteristic  appearances,  micro-organisms 
which,  in  their  individual  form,  closely  resemble  one  another,  or 
are  even  identical.  The  same  process,  with  slight  modification,  is 
also  employed  in  the  examination  of  air,  soil,  and  water,  which  will 
be  referred  to  later. 

The  preparation 
of  plate-cultivations, 
therefore,  must  be 
described  in  every  de- 
tail :  and  to  take  an 
example,  we  will  sup- 
pose that  a  series  of 
plates  is  to  be  pre- 
pared from  a  test- 
cube- cultivation. 

Arrangement      of 

Levelling  Apparatus. — In  order  to  spread  out  the  liquid  jelly 
evenly  on  the  surface  of  a  glass  plate,  and  hasten  its  solidifica- 
tion, it  is  necessary  to  place  the  glass  plate  upon  a  level  and 
cool  surface.  This  is  obtained  in  the  following  manner :  Place  a 
large  shallow  glass  dish  upon  a  tripod  stand,  and  fill  it  to  the  brim 
with  cold  water ;  carefully  cover  the  dish  with  a  slab  of  plate-glass, 
or  a  pane  of  window-glass,  and  level  it  by  placing  the  spirit-level  in 
the  centre  and  adjusting  the  screws  of  the  tripod.  Substitute  for 
the  spirit-level  a  piece  of  filter-paper  the  size  of  the  glass  plates  to 
be  employed,  and  cover  it  with  a  shallow  bell-glass. 

Sterilisation  of  Glass  Plates. — The  glass  plates  are  sterilised  in 
an  iron  box  placed  in  the  hot-air  steriliser,  at  150°  C.,  from  one  to 
two  hours.  As  these  plates  are  used  also  for  other  purposes,  a 
quantity  ready  sterilised  should  always  be  kept  in  the  box. 

Preparation  of  Damp  Chambers. — The  damp  chambers  for  the 
reception  of  the  inoculated  plates  are  prepared  thus  :  Thoroughly 


FIG.  36. — LEVELLING  APPARATUS. 


108 


BACTERIOLOGY. 


cleanse  and  wash  out  with  1  in  20  carbolic  acid  a  shallow  glass  dish 
and  bell.     Cut  a  piece  of  filter-paper  to  line  the  bottom  of  the  glass 
dish,  and  moisten  it  with  the  same  solution. 
Method  of  Inoculating  the  Test-tubes. — In 
a  glass  beaker  or  an  ordinary  glass  tumbler, 
with  a  pad    of    cotton-wool    at  the  bottom, 
place  the    tube    containing   the   cultivation, 
the  three  tubes  to  be  inoculated,  three  glass 
rods  which  have  been  sterilised   by  heating 
in    the    flame  of    a    Bunsen  burner,   and    a 
thermometer.     Provide  a   strip   of  paper,   a 
large  label,  a  pencil,  a  pair  of  forceps,  and 
inoculating  needles.     All    is    now    ready  at 
hand  to  commence  the  inoculation  of  the  tubes. 
Liquefy  the  gelatine  in  the  three  tubes  by  placing  them  in  a 
beaker  containing  water  at  30°  C.,  or  by  gently  warming  them  in 
the  flame  of  the  Bunsen  burner.     Keep  the  tubes,  both  before  and 
after  the  inoculation,  in  the  warm  water,  to  maintain  the  gelatine 
in  a  state  of  liquefaction.     Hold  the  tube  containing  the  cultivation 


FIG.  37. — IRON    Box 
GLASS  PLATES. 


FOR 


FIG.  38.— METHOD  OF  INOCULATING  TEST-TUBES  IN  THE  PREPARATION 
OF  PLATE-CULTIVATIONS. 

and  a  tube  of  the  liquefied  gelatine  as  nearly  horizontal  as  possible 
between  the  thumb  and  index  finger  of  the  left  hand.  With  the  index 
finger  and  thumb  of  the  right  hand  loosen  the  plugs  of  the  tubes. 
Take  the  looped  platinum  needle  in  the  right  hand  and  hold  it  like 
a  pen.  Remove  the  plug  from  the  culture- tube  by  using  the  fourth 
and  fifth  fingers  of  the  right  hand  as  forceps,  and  place  it  between 


DESCRIPTION    OF    PLATE    III. 
Plate-cultivation. 

This  represents  the  appearance  of  a  plate-cultivation  of  the  comma-bacillus 
of  Cholera  nostras,  when  it  is  examined  over  a  slab  of  blackened  plate-glass. 
The  drawing  was  made  from  a  typical  result  of  thinning  out  the  colonies  by 
the  process  of  plate-cultivation.  At  this  stage  they  were  completely  isolated 
one  from  the  other ;  but  later  they  became  confluent,  and  produced  complete 
liquefaction  of  the  gelatine. 


Plate  IE. 


fc 

o 

f-H 

H 

t 

i— i 
H 

H 
P 
O 


NUTRIENT   MEDIA    AND   METHODS   OF   CULTIVATION.  109 

the  fourth  and  fifth  fingers  of  the  left.  Remove  the  plug  of  the 
other  tube  in  the  same  way,  placing  it  between  the  third  and  fourth 
fingers  of  the  left  hand.  With  the  needle  take  up  a  droplet  of  the 
cultivation  and  stir  it  round  in  the  liquefied  jelly.  Replace  the  plugs, 
and  set  aside  the  cultivation.  Hold  the  freshly  inoculated  tube  be- 
tween the  index  finger  and  thumb  of  either  hand,  almost  horizontally, 
then  raise  it  to  the  vertical,  so  that  the  liquid  gelatine  gently  flows 
back.  By  repeating  this  motion  and  rolling  the  tube  between  the 
fingers  and  thumbs  the  micro-organisms  which  have  been  introduced 
are  distributed  throughout  the  gelatine.  Any  violent  shaking,  and 
consequent  formation  of  bubbles,  must  be  carefully  avoided.  From 
the  inoculated  tube,  in  the  same  manner  inoculate  a  fresh  tube  of 
liquefied  gelatine,  introducing  into  it  three  droplets  with  a  sterilised 
needle.  After  tilting  and  rolling  this  tube,  as  in  the  previous  case, 
the  same  process  is  repeated  with  a  third  tube,  which  is  inoculated 
from  the  second  tube.  This  last  tube  must  be  inoculated  in  different 
ways,  according  to  experience,  for  different  micro-organisms.  Some- 
times a  sufficient  separation  of  the  micro-organisms  is  attained  by 
inoculating  the  last  tube  with  a  straight,  instead  of  a  looped,  needle, 
dipping  it  from  the  one  into  the  other  from  three  to  five  times. 

The  next  process  consists  in  pouring  out  the  gelatine  on  glass 
plates  and  allowing  it  to  solidify. 

Preparation  of  the  Gelatine- Plates. — The  directions  to  be  observed 
in  pouring  out  the  gelatine  are  as  follows  : — 

Place  the  box  containing  sterilised  plates  horizontally,  and  so 
that  the  cover  projects  beyond  the  edge  of  the  table ;  remove  the 
cover,  and  withdraw  a  plate  with  sterilised  forceps  ;  hold  it  between 
the  finger  and  thumb  by  opposite  margins,  rapidly  transfer  it  to 
the  filter-paper  under  the  bell-glass,  and  quickly  replace  the  cover 
of  the  box.  On  removing  the  plug  from  the  tube  which  was  first 
inoculated,  an  assistant  raises  the  bell-glass,  and  the  contents  of  the 
tube  are  poured  on  to  the  plate ;  with  a  glass  rod  the  gelatine  must 
be  then  rapidly  spread  out  in  an  even  layer  within  about  half  an 
inch  of  the  margin  of  the  plate.  The  assistant  replaces  the  bell- 
glass,  and  the  gelatine  is  left  to  set.  Meanwhile  a  glass  bench  or 
metallic  shelf  is  placed  in  the  damp  chamber,  ready  for  the  reception 
of  the  plate-cultivation,  and  when  the  gelatine  is  quite  solid  the 
plate  is  quickly  transferred  from  under  the  bell-glass  to  the  damp 
chamber ;  precisely  the  same  process  is  repeated  with  the  other 
tubes,  and  the  damp  chamber,  labelled  with  the  details  of  the 
experiment,  is  set  aside  for  the  colonies  to  develop.  Not  only  plate- 
cultures  should  be  carefully  labelled  with  date  and  description,  but 


110  BACTERIOLOGY. 

the  same  remark  applies  equally  to  all  preparations — tube- cultures, 
potato-cultures,  drop-cultures,  etc. 

Corresponding  with  the  fractional  cultivation  of  the  micro- 
organisms obtained  in  this  manner,  the  colonies  will  be  found  to 
develop  in  the  course  of  a  day  or  two,  the  time  varying  with  the 
temperature  of  the  room.  The  lower  plate  will  contain  a  countless 
number  of  colonies  which,  if  the  micro-organism  liquefies  gelatine, 
speedily  commingle,  and  produce,  in  a  very  short  time,  a  complete 
liquefaction  of  the  whole  of  the  gelatine.  On  the  middle  plate  the 
colonies  will  also  be  very  numerous,  but  retain  their  isolated  position 
for  a  longer  time  ;  while  on  the  uppermost  plate  the  colonies  are 
completely  isolated  from  one  another,  with  an  appreciable  surface  of 
gelatine  intervening. 

Examination  of  Plate- cultivations. — The  macroscopical  appear- 
ances of  the  colonies  are  best  studied  by  placing  the  plate  on  a 


FIG.  39. — DAMP  CHAMBER  CONTAINING  PLATE-CULTIVATIONS. 

slab  of  blackened  glass,  or  on  a  porcelain  slab  if  the  colonies  are 
coloured. 

To  examine  the  microscopical  appearances,  a  selected  plate  is 
placed  upon  the  stage  of  the  microscope.  The  smallest  diaphragm 
is  employed,  and  the  appearances  studied  principally  with  a  low 
power.  These  appearances  should  be  carefully  noted,  and  a 
sketch  or  photograph  of  the  colony  made.  The  morphological 
characteristics  of  the  micro-organisms  of  which  the  colony  is  formed 
can  be  examined  in  the  following  way  :  A  small  looped  or 
hooked  platinum  needle  is  held  like  a  pen.  and  the  hand  steadied 
by  resting  the  little  finger  on  the  stage  of  the  microscope.  The 
extremity  of  the  needle  is  steadily  directed  to  the  space  between 
the  lens  and  the  gelatine  without  touching  the  latter,  until,  011 
looking  through  the  microscope,  it  can  be  seen  in  the  field,  above 
or  by  the  side  of  the  colony  under  examination.  The  needle 
is  then  dipped  into  the  colony,  steadily  raised,  and  withdrawn. 
Without  removing  the  eye  from  the  microscope  this  manipulation 
can  be  seen  to  be  successful  by  the  colony  being  disorganised  or 


NUTRIENT   MEDIA    AND   METHODS   OF   CULTIVATION. 


Ill 


completely  removed  from  the  gelatine.  It  is,  however,  not  easy  to 
lu«  successful  at  first,  but  with  practice  this  can  be  accomplished 
with  rapidity  and  precision.  A  preparation  is  then  made  by  rubbing 


FIG.  40.— PASTEUR'S  LARGE  INCUBATOR. 


the  extremity  of  the  needle  in  a  droplet  of  water  on  a  slide,  covering 
with  a  cover-glass,  and  examining  in  the  fresh  state,  or  by  spreading 
out  the  droplet  on  a  cover-glass,  drying,  passing  three  times  through 
the  flame,  and  staining  with  a  drop  of  fuchsine  or  gentian  violet. 
Inoculations  should  be  made  in  test-tubes  of  nutrient  gelatine 


OF   THF. 

UNIVERSITY 


112  BACTERIOLOGY. 

and  agar-agar,  from  the  micro-organisms  transferred  to  the  cover- 
glass  before  it  is  dried  and  stained,  from  any  remnants  of  the  colony 
which  was  examined,  or  from  other  colonies  bearing  exactly  similar 
appearances.  In  this  way  pure  cultivations  are  established,  and 
the  rnacroscopical  appearances  of  the  growth  in  test-tubes  can  be 
obtained.  The  plates  should  be  replaced  in  the  damp  chamber 
as  soon  as  possible ;  drying  of  the  gelatine,  or  contamination  with 
micro-organisms  gravitating  from  the  air  during  their  exposure, 

may  spoil  them  for  subsequent  examination. 
A  much  simpler  method  of  plate-cultivation 

is  to  dispense  with  the  levelling  apparatus,  and 

pour  the  liquefied  ielly  into  shallow,  flat  dishes. 
FIG.  41.— PETRI'S  J     J 

j)ISH<  -They  take  up  much  less  room,   and  in   many 

ways  are  more  convenient  (Fig.  41). 

Nutrient  agar-agar  can  also  be  employed  for  the  preparation 
of  plate-cultivations,  but  it  is  much  more  difficult  to  obtain  satis- 
factory results.  The  test-tubes  of  nutrient  agar-agar  must  be 
placed  in  a  beaker  with  water  and  heated  until  the  agar-agar 
is  completely  liquefied.  The  gas  is  then  turned  down,  and  the 
temperature  of  the  water  allowed  to  fall  until  the  thermometer 
stands  just  above  50°  C.  The  water  must  be  maintained  at  this 
temperature,  and  the  test-tubes  must  be  in  turn  rapidly  inoculated 
and  poured  out  upon  the  glass  plates,  or  better  still,  into  glass 
dishes,  as  already  described. 

A  very  much  simpler  plan  is  to  liquefy  the  agar,  pour  it  into 


FIG.  42. — GLASS  BENCHES  AND  SLIDES. 

a  shallow  dish,  and  allow  it  to  solidify.  The  culture  material  is 
thinned  out  in  sterilised  broth,  and  a  few  drops  are  spread  out  over 
the  surface  of  the  agar.  The  dishes  are  then  placed  in  the  incubator 
at  37°  C. 

Glass  plates  may  also  be  employed  in  a  much  simpler  way. 
The  nutrient  jelly  is  liquefied,  poured  out,  and  allowed  to  set.  A 
needle  charged  with  the  material  to  be  inoculated  is  then  drawn 
in  lines  over  the  surface  of  the  jelly.  This  method  is  of  use  for 
inoculating  different  organisms  side  by  side,  and  watching  the  effect 
of  one  upon  the  other,  or  a  micro-organism  in  this  way  may  be 


NUTRIENT   MEDIA    AND   METHODS   OF   CULTIVATION.  1 13 

sown  upon  the  gelatine  which  has  been  already  altered  by  the 
growth  of  another  micro-organism  ;  the  change  produced  in  the 
gelatine,  as  in  the  case  of  the  Bacillus  pyocyaneus,  extending  far 
beyond  the  limits  of  the  growth  itself. 

Nutrient  jelly  may  also  be  spread  out  on  sterilised  glass  slides, 
which  after  inoculation  are  placed  in  damp  chambers  for  the  growths 
to  develop. 

Esmareh's  Roll-cultures. — Esmarch  introduced  a  modification 
of  the  method  of  plate- cultivation  which  may  sometimes  be  used  with 
advantage.  The  ordinary  test-tubes  may  be  employed,  or  tubes 
considerably  larger  in  size. 

After  the  liquid  jelly  has  been  inoculated  in  the  tube,  instead 
of  pouring  it  out  on  to  a  glass  plate  or  into  a  dish,  the  cotton-wool 
plug  is  replaced,  and  an  india-rubber  cap  fitted  over  the  mouth  of 
the  tube. 

The  tube  is  then  placed  horizontally  on  a  block  of  ice  or 
in  a  vessel  containing  iced  water.  The  neck  of  the  tube  is  steadied 
with  the  left  hand,  and  the  tube  turned  round  and  round  with  the 
right  hand.  In  a  very  short  time  the  gelatine  sets,  and  the  tube 
is  lined  inside  with  a  thin  coating.  There  is  far  less  danger  of 
contamination,  and  the  cultures  are  in  a  much  more  convenient 
form  when  circumstances  render  it  necessary  to  move  them. 

(c)  PREPARATION  AND  EMPLOYMENT  OF  SOLIDIFIED  BLOOD  SERUM. 

Solid  Blood  Serum.— The  tubercle-bacillus,  the  bacilli  of 
glanders  and  of  diphtheria,  and  many  other  micro-organisms,  thrive 
well  when  cultivated  on  solid  blood  serum.  This  medium  has  the 
additional  advantage  of  remaining  solid  at  all  temperatures.  The 
technique  required  for  its  preparation  and  sterilisation  is  as  follows  : 
Several  cylindrical  vessels,  about  20  cm.  high,  are  thoroughly  washed 
with  carbolic  acid  (1  in  20),  and  then  with  alcohol,  and  finally 
rinsed  out  with  ether.  The  ether  is  allowed  to  evaporate,  and  the 
vessels  are  then  ready  for  use.  The  skin  of  the  animal  selected — 
calf,  sheep,  or  horse — is  washed  with  carbolic  at  the  seat  of  operation, 
and  the  bleeding  performed  with  a  sterilised  knife  or  a  trocar  and 
canuula.  The  first  jet  of  blood  from  the  vein  is  rejected,  and  that 
which  follows  is  allowed  to  flow  into  the  vessels  until  they  are  almost 
full.  The  ground-glass  stoppers,  greased  with  vaseline,  are  replaced, 
and  the  vessels  set  aside  in  ice,  as  quickly  as  possible,  for  from 
twenty-four  to  thirty  hours.  By  that  time  the  separation  of  the 
clot  is  completed,  and  the  clear  serum  can  then  be  transferred  to 

8 


114 


BACTERIOLOGY. 


plugged  sterilised  test-tubes.  These  should  be  filled,  with  a  sterilised 
pipette,  to  about  one-third  of  their  capacity. 

Formerly  the  tubes  were  sterilised  by  Tyndall's  process  of  dis- 
continuous sterilisation.  The  tubes  were  placed  in  Koch's  serum 
steriliser,  with  the  temperature  maintained  for  an  hour  or  more  at 
56°  C.,  and  this  was  repeated  for  six  successive  days,  the  temperature 
on  the  last  day  being  gradually  raised  to  60°  C.  This  completed  the 
sterilisation,  and  to  solidify  the  serum  the  tubes  were  arranged  in 
the  inspissator  at  the  angle  required,  and  the  temperature  was  kept 

between  65°  C.  and  68°  C. 
Directly  solidification  took 
place  the  tubes  were  removed. 
The  new  process  is  much 
less  tedious,  and  consists  in 
taking  every  possible  pre- 
caution to  obtain  the  blood 
without  contamination  by  bac- 
teria in  the  air  or  in  the 
vessels  employed.  There  is 
then  no  need  to  sterilise  the 
serum,  and  it  can  be  coagu- 
lated immediately.  The  tubes 
are  tested  by  placing  them 
in  an  incubator  at  37°  C.  for 
a  week,  and  if  any  show 
signs  of  contamination  they 
are  discarded,  and  the  rest 
can  be  used  or  kept  in  stock. 

The    serum    should    then 

present  the  character  of  being  hard,  solid,  of  a  pale  straw  colour, 
and  transparent.  A  little  liquid  collects  at  the  lowest  point,  and 
the  serum  is  sometimes  milky  in  appearance  at -its  thickest  part. 

Loffler's  Blood  Serum  is  prepared  by  mixing  two- thirds  of  fresh 
serum  with  one-third  of  broth,  prepared  in  the  usual  way  but  with 
the  addition  of  1  per  cent,  grape-sugar.  The  mixture  is  decanted 
into  test-tubes,  avoiding  the  formation  of  air- bubbles,  and  it  is  then 
coagulated  in  the  usual  way.  The  serum  may  be  employed  not  only 
in  test-tubes,  but  also  in  small  flasks,  glass  capsules,  or  other  vessels, 
all  of  which  must  be  cleansed  and  sterilised. 

Hydrocele  fluid  and  other  serous  effusions  may  be  prepared  in 
the  same  manner.  (Gelatine  may  be  added  to  the  serum  in  the  pro-- 
portion of  5  per  cent. 


FIG.  43. — KOCH'S  SERUM  STERILISER. 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION.  115 

Inoculation  of  the  Tubes. — A  small  portion  of  a  culture  or  of  the 
material  to  be  inoculated  is  taken  up  with  a  sterilised  platinum 
needle,  and  traced  over  the  sloping  surface  of  the  serum ;  or  a 
fragment  of  tissue,  such  as  diphtheritic  membrane  or  tubercle,  may 
be  introduced  into  the  tube  and  rubbed  gently  over  the  serum  so  as 
not  to  break  the  surface. 


FIG.  44. — HUEPPE'S  SERUM  INSPISSATOR. 

(D)  PREPARATION  AND  EMPLOYMENT  OF  STERILISED  POTATO. 

Potato-cultivations. — Sterilised  potatoes  form  an  excellent 
medium  for  the  cultivation  of  many  micro-organisms,  more  especially 
the  chromogenic  species.  Potato -cultivations  also  give  in  some  cases 
very  characteristic  appearances,  which  are  of  value  in  distinguishing 
bacteria  which  possess  morphological  resemblances. 

Preparation  of  Sterilised  Potatoes. — Potatoes,  preferably  smooth  - 
skinned,  which  are  free  from  "eyes"  and  rotten  spots,  should  be 
selected.  If  they  cannot  be  obtained  without  eyes  and  spots,  these 
must  be  carefully  picked  out  with  the  point  of  a  knife.  The  potatoes 
are  well  scrubbed  with  a  stiff  brush,  and  allowed  to  soak  in  1  in  20 
carbolic  for  a  few  minutes.  They  are  then  transferred  to  the  potato- 


116  BACTERIOLOGY . 

receiver,  and  steamed  in  the  steam  steriliser  for  twenty  minutes  to 
half  an  hour,  the  time  varying  according  to  the  size  of  the  potatoes. 
When  cooked,  the  potato-receiver  is  withdrawn  and  left  to  cool,  the 
potatoes  being  retained  in  it  until  required  for  use. 

Damp  chambers  are  prepared  ready  for  the  potatoes,  the  vessels 
being  cleansed  and  washed  with  carbolic  as  described  for  plate- 
cultivations.  Small  glass  dishes  of  the  same  pattern  as  the  large 
ones  may  be  employed  for  single  halves  of  potatoes.  Potato-knives 
and  scalpels,  which  have  been  sterilised  in  an  iron  box  by  heating 
them  in  the  hot-air  steriliser  at  151°  C.  for  one  hour,  should 
be  ready  to  hand.  Knives  sterilised  by  heating  them  in  the  name 
of  a  Bunsen  burner  should  afterwards  be  placed  upon  a  sterilised 
glass  plate  and  covered  with  a  bell-glass.  It  must  not  be  forgotten, 


FIG.  45. — Box  FOR  STERILISING  INSTRUMENTS. 

however,  that  heating  the  blades  in  the  flame  destroys  the  temper 
of  the  steel,  and  therefore  knives  and  other  instruments  should 
preferably  be  sterilised  in  the  hot-air  steriliser,  enclosed  in  an  iron 
box,  or  simply  enveloped  in  cotton-wool. 

Inoculation  of  Potatoes. — The  coat-sleeves  should  be  turned  back, 
and  the  hands,  after  thorough  washing  with  good  lathering  soap, 
be  dipped  in  1  in  40  carbolic.  An  assistant  opens  the  potato- 
receiver,  and  a  potato  is  selected  and  held  between  the  thumb  and 
index  linger  of  the  left  hand.  With  the  knife  held  in  the  right 
hand,  the  potato  is  almost  completely  divided  in  the  direction 
which  will  give  the  largest  surface.  The  assistant  raises  the  cover 
of  the  damp  chamber,  and  the  potato  is  introduced,  and  while  the 
knife  is  withdrawn,  allowed  to  fall  apart.  The  cover  is  quickly 
replaced,  and  another  potato  treated  in  the  same  way,  is  placed 
in  the  same  damp  chamber.  The  four  halves  are  then  quite  ready 
for  inoculation.  As  an  extra  precaution,  the  left  hand  is  again 
dipped  in  carbolic,  and  one  half  of  a  potato  is  taken  up  between  the 
tips  of  the  thumb  and  index  finger,  care  being  taken  to  avoid 
touching  the  cut  surface.  Holding  it  with  its  cut  surface  vertical, 


NUTRIENT    MEDIA   AND   METHODS    OF   CULTIVATION.  117 

a  small  portion  of  the  substance  to  be  inoculated  is  placed  on  the 
centre  with  a  sterilised  platinum  needle.  With  a  sterilised  scalpel 
the  inoculated  substance  is  rapidly  spread  over  the  surface  of  the 
potato  with  the  flat  of  the  blade,  to  within  a  quarter  of  an  inch 
of  the  margin,  and  the  potato  is  then  as  quickly  as  possible  replaced 
in  the  damp  chamber.  With  another  sterilised  scalpel  a  small 
portion  of  the  potato  from  the  inoculated  surface  of  the  first  half 
is  in  the  same  way  spread  over  the  surface  of  the  second  half,  thus 
thinning  out  the  bacteria  as  in  plate-cultivations.  Exactly  the 
same  is  repeated  with  a  third  potato,  and  even  a  fourth,  so  that 
a  still  further  thinning  out  or  fractional  cultivation  of  the  micro- 
organisms may  be  obtained.  In  some  cases  it  is  necessary  to  place 
the  cultures  in  an  incubator  (Fig.  40) ;  others  grow  very  well  at  the 


FIG.  46. — DAMP  CHAMBER  FOR  POTATO-CULTIVATIONS. 

temperature  of  the  room.  As  in  plate-cultivations,  the  potato  may 
also  be  inoculated  by  simply  streaking  it  in  lines  with  a  needle 
charged  with  the  material  to  be  cultivated. 

Potato  in  Test-tubes. — Large  surfaces  of  potato  are  employed  when 
we  wish  to  obtain  cultures  of  micro-organisms  in  considerable  quanti- 
ties, as  in  the  examination  of  the  products  of  chromogenic  bacteria  ; 
but  under  ordinary  circumstances  potato  is  employed  in  test-tubes. 
The  central  portions  of  raw  potatoes  are  cut  out  in  cylindrical  pieces 
with  a  cork-borer.  These  are  divided  obliquely  in  their  whole 
length,  and  each  half  is  placed  in  a  test-tube.  The  test-tubes  are 
plugged  with  cotton-wool,  and  then  steam  in  the  steam-steriliser 
for  twenty  minutes.  The  sloping  surface  is  inoculated  in  the  same 
way  as  obliquely  solidified  jelly,  and  the  advantages  are  great.  The 
cultures  are  obtained  in  a  more  convenient  form,  and  there  is  less 
danger  of  contamination. 

Potato-paste  may  be  employed  when  it  is  desirable  to  obtain  an 
extensive  growth  of  certain  bacteria.  The  potatoes  are  boiled  for 
an  hour,  and  the  floury  centre  squeezed  out  of  the  skins.  This 
is  then  mashed  up  with  sufficient  sterilised  water  to  produce  a  thick 


118  BACTERIOLOGY. 

paste,  and  is  heated  in  the  steam  steriliser  for  half  an  hour  for  three 
successive  clays. 

(E)  PREPARATION  AND  EMPLOYMENT  OF  BREAD-PASTE,  VEGETABLES, 
FRUIT,  WHITE  OF  EGG. 

Some  micro-organisms,  more  especially  mould  fungi,  grow  very 
well  on  bread-paste.  This  is  prepared  by  removing  the  crust  from 
slices  of  bread  and  drying  them  in  the  oven.  They  are  then 
broken  up,  and  reduced  to  a  fine  powder  with  a  pestle  and  rnortar. 
Small,  carefully  cleansed,  conical,  or  globe-shaped  flasks  are  plugged 
with  cotton-wool  and  sterilised  in  the  oven.  When  cool,  a  small 
quantity  of  the  powder  is  placed  in  them,  and  sterilised  water  added 
in  the  proportion  of  one  part  to  every  four  of  the  powder.  The 
paste  is  sterilised  by  steaming  in  the  steriliser  at  100°  C.  for  half  an 
hour  for  three  successive  days.  The  flasks  can  be  reversed,  and  may 
be  inoculated  with  a  platinum  needle. 

Boiled  carrots  and  other  vegetables,  and  various  kinds  of  stewed 
fruit,  are  also  occasionally  employed  for  the  cultivation  of  bacteria. 
The  sterilisation  of  these  media  must  be  carried  out  on  the  principles 
already  explained. 

White  of  egg  may  be  solidified  in  shallow  glass  dishes,  in  the 
steam  steriliser.  After  inoculation  the  dishes  should  be  placed 
in  a  damp  chamber. 

LIQUID  MEDIA. 

(F)  PREPARATION  OF  STERILISED  BROTH,  LIQUID  BLOOD  SERUM, 
URINE,  MILK,  VEGETABLE  INFUSIONS,  AND  ARTIFICIAL  NOURISH- 
ING LIQUIDS. 

Nutrient  liquids  are  still  largely  employed.  For  inoculation  ex- 
periments when  the  presence  of  gelatine  is  undesirable,  for  studying 
the  physiology  and  chemistry  of  bacteria,  and  when  for  any  object 
a  rapid  growth  of  micro-organisms  is  necessary,  the  employment 
of  liquid  media  is  not  only  advisable,  but  absolutely  necessary. 
Liquid  media  comprise  two  distinct  groups — natural  and  artificial. 
Natural  media  include  meat  broth,  blood,  urine,  milk,  and  vegetable 
infusions ;  artificial  media  are  solutions  composed  from  a  chemical 
formula  representing  essential  food  constituents. 

Broth  may  be  made  from  beef,  pork,  chicken,  or  fish  in  the 
manner  which  has  been  described  for  the  preparation  of  nutrient 
gelatine,  simply  with  omission  of  the  gelatine.  After  the  process 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION.  119 

of  neutralisation  with  carbonate  of  soda  solution,  the  flask  of  broth 
is  placed  in\  the  steam  steriliser  for  half  an  hour  at  100°  C.  A 
clear  liquid  results  on  nitration  which  is  transferred  to  plugged 
sterilised  flasks  or  test-tubes,  and  sterilisation  effected  by  exposing 
the*n  in  the  steam  steriliser  for  half  an  hour  at  100°  C.  for  two 


FIG.  47. — APPARATUS  FOE  STERILISATION  BY  STEAM  UNDER  PRESSURE. 

or  three  successive  days,  or]  by;  I  using  the  apparatus  for  sterilising 
by  steam  under  pressure.  For  some  bacteria  a  more  suitable 
cultivating  medium  is  obtained  by  the  addition  of  glycerine  or 
grape-sugar. 

Liquid  Blood  Serum. — The  preparation  of  blood  serum  has 
already  been  described.  '  It  may  be  required  for  cultivations  before 
the  final  treatment  by  which  it  is  solidified,  for  example,  in  the 
method  of  drop-cultivation,  and  may  be  used  with  the  addition  of 
glycerine  or  grape-sugar.  Hydrocele  fluid,  peritonitic  arid  pleuritic 


120  BACTERIOLOGY. 

effusions,  can  also  be  employed  after  sterilisation  in  the  steam 
steriliser.  The  fluid  should  be  withdrawn  with  a  sterilised  trocar 
and  cannula,  and  received  into  plugged  sterilised  flasks. 

Urine. — -In  order  to  obtain  urine  free  from  micro-organisms  the 
following  precautions  must  be  observed  :  The  orifice  of  the  urethra 
must  be  thoroughly  cleansed  with  weak  carbolic.  The  first  jet  of 
urine  should  be  rejected,  and  the  rest  received  into  sterilised  vessels, 
which  must  be  quickly  closed  with  sterilised  plugs.  If  these  pre- 
cautions be  not  attended  to,  the  urine  must  be  rendered  sterile  by 
the  means  described  for  the  sterilisation  of  broth. 

Milk. — If  milk  has  been  drawn  into  sterile  flasks,  after  thoroughly 
cleansing  and  disinfecting  the  teats  and  hands,  it  may  be  kept 
without  change.  If  procured  without  these  precautions,  it  must 
be  steamed  in  the  steriliser  for  half  an  hour  for  five  successive 
days. 

Vegetable  and  other  Infusions. — Infusions  of  hay,  cucumber, 
and  turnip  are  used  for  special  purposes,  and  more  rarely  decoctions 
of  plums,  raisins,  malt,  and  horse-dung.  They  are  mostly  prepared 
by  boiling  with  distilled  water,  after  maceration  for  several  hours. 
The  filtrate  is  received  into  sterile  flasks  and  sterilised  in  the  usual 
way  in  the  steam  steriliser. 

Artificial  Fluids. — Pasteur's  solution  is  prepared  by  mixing  the 
ingredients  in  the  following  proportions  : — 

Distilled  water .100 

Pure  cane-sugar   .         .         .         .         .         .         10 

Ammonium  tartrate     .....  1 

Ash  of  yeast '075 

Mayer's  modification  of  the  nourishing  fluid  employed  by  Colin  is 
as  follows : — 

Distilled  water 20 

Ammonium  tartrate "2 

Phosphate  of  potassium          .         .         .         .  "1 

Sulphate  of  magnesium                   ...  "1 

Tribasic  calcium  phosphate    ....  '01 

Drop- cultures. — This  method  of  cultivation  is  a  particularly 
instructive  one.  It  enables  us  to  study  many  of  the  changes  which 
take,  place  during  the  life  history  of  micro-organisms.  This  is 
illustrated,  for  example,  in  a  drop-culture  of  the  anthrax  bacillus, 
in  which  we  can  watch  the  gradual  growth  of  a  single  bacillus  into 


NUTRIENT   MEDIA   AND   METHODS    OF   CULTIVATION. 


121 


a  long  filament,  and  the  subsequent  development  of  bright  oval 
spores.  It  is  necessary  carefully  to  observe  the  minutest  details 
in  order  to  maintain  the  cultivation  pure.  An  excavated  slide  is 
thoroughly  cleaned,  and  then  sterilised  by  being  held  with  the 
cupped  side  downwards  in  the  flame  of  the  Bunsen  burner.  A  ring 
of  vaseline  is  painted  round  the  excavation,  and  the  slide  is  then 
placed  under  a  glass  bell.  Meanwhile  a  carefully  cleansed  cover- 
glass  is  also  sterilised  by  passing  it  through  the  flame,  and  should 
be  deposited  on  a  sterilised  glass  plate.  With  a  sterilised  looped 
needle,  a  drop  of  sterile  broth  is  transferred  to  the  cover-glass, 
and  this  is  inoculated  by  touching  it  with  another  sterilised  needle 
charged  with  the  material  to  be  examined,  without  disturbing 
the  form  of  the  drop.  It  is  quite  sufficient  just  to  touch  the  drop 


FIG.  48. — DROP  CULTIVATION. 
(a)  Drop  of  broth ;    (6)  layer  of  vaseline. 

instead  of  transferring  a  visible  quantity  of  blood,  juice,  or  growth, 
as  the  case  may  be.  The  slide  is  then  inverted  and  placed  over  the 
cover-glass,  so  that  the  drop  will  come  exactly  in  the  centre  of  the 
excavation,  and  is  gently  pressed  down.  On  turning  the  slide  over 
again  the  cover-glass  adheres,  and  an  additional  layer  of  vaseline 
is  painted  round  the  edges  of  the  cover-glass  itself.  The  slide  must 
be  labelled,  and  if  necessary,  placed  in  the  incubator,  and  the  results 
watched  from  time  to  time.  Instead  of  broth,  liquid  blood  serum 
may  be  employed  in  this  form  of  cultivation.  If  it  is  required 
to  preserve  the  drop-cultivation  as  a  microscopic  preparation,  the 
cover-glass  is  gently  lifted  off  and  allowed  to  dry.  Any  vaseline 
adhering  to  the  cover-glass  should  be  wiped  off,  and  the  cover-glass 
can  then  be  passed  through  the  flame  and  stained  in  the  usual  manner. 
Moist  Cells. — Unless  drop-cultures  are  very  carefully  prepared 


122  BACTERIOLOGY. 

they  are  liable  to  dry  up,  if  kept  for  examination  for  several  days. 
Many  therefore  prefer  employing  a  moist  cell,  of  which  there  are 
several  different  forms  in  use. 

The  drop-culture  slide  may  be  converted  into  a  moist  cell 
by  having  a  deep  groove  cut  round  the  circumference  of  the  con- 
cavity. This  groove  is  filled  with  sterilised  water  by  means  of  a 
pipette.  A  ring  of  vaseline  is  painted  with  the  camel's-hair  brush 
outside  the  groove,  and  the  cover-glass,  with  the  drop -cultivation, 
is  inverted  and  placed  over  the  concavity.  This  form  is  very  useful, 
as  the  slide  can  be  easily  cleansed  and  effectually  sterilised  by 
holding  it  in  the  flame  of  the  Bunsen  burner. 

A    very   simple   form    of   moist    cell   recommended   by    Schafer 


FIG.  49. — SIMPLE  METHOD  OF  FOKMING  A  MOIST  CELL. 

may  be  used  in  some  cases,  but  possesses  the  disadvantage  of  not 
admitting  of  sterilisation  by  heat.  A  small  piece  of  putty  or 
modelling  wax  is  rolled  into  a  cord  about  two  inches  long  and  J  inch 
thick.  By  uniting  the  ends  a  ring  is  formed,  which  is  placed  on  the 
middle  of  a  clean  glass  slide.  A  drop  of  water  is  placed  in  the 
centre  of  the  ring,  and  the  cell  roofed  in  by  applying  a  cover-glass. 

A  cell  somewhat  similar  in  form,  which  has  the  advantage  of 
permitting  of  thorough  cleansing,  may  be  constructed  by  cementing 
a  glass  ring  with  flat  surfaces  to  an  ordinary  slide.  Vaseline  is 
applied  with  a  camel's-hair  brush  to  the  upper  surface  of  the  ring, 
and  one  or  two  drops  of  water  placed  with  a  pipette  at  the  bottom 
of  the  cell.  The  cover-glass,  with  the  preparation,  is  then  inverted 
over  the  cell  and  gently  pressed  down  upon  the  glass  ring.  The 
vaseline  renders  the  cell  air-tight,  and,  to  a  certain  extent,  fixes 
the  cover-glass  to  the  ring. 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


123 


Warm  Stages. — To  apply  warmth  while  a  preparation  is  under 
continuous  observation,  we  must  either  place  the  microscope  bodily 


FIG.  50.— WARM  STAGE. 


within  a  special  incubator,  with  the  eye-piece  protruding  through  an 
opening,  or  we  must  employ  some  means  of  applying  heat  directly 
to  the  preparation. 

A  simple  warm  stage  may  be  made  of  an  oblong  copper  plate, 


FIG.  51. — WARM  STAGE  SHOWN  IN  OPERATION. 

two  inches  long  by  one  inch  wide,  from  one  side  of  which  a  rod  of 
the  same  material  projects.     The  plate  has  a  round  aperture  in  the 


124  BACTERIOLOGY. 

middle,  half  an  inch  in  diameter,  and  is  fastened  to  an  ordinary 
slide  with  sealing-wax.  The  drop  to  be  examined  is  placed  on  a 
large-sized  cover-glass  and  covered  with  a  smaller  one.  Olive  oil 
or  vaseline  is  painted  round  the  edge  of  the  smaller  cover-glass  to 
prevent  evaporation,  and  the  preparation  is  placed  over  the  aperture 
in  the  plate. 

The  slide  bearing  the  copper  plate  is  clamped  to  the  stage  of 


FIG.  52.— ISRAEL'S  WARMING  APPARATUS  IN  OPERATION. 

the  microscope.  The  flame  of  a  spirit-lamp  is  applied  to  the 
extremity  of  the  rod,  and  the  heat  is  conducted  to  the  plate  and 
thence  transmitted  to  the  specimen.  In  order  that  the  temperature 
of  the  copper  plate  may  be  approximately  that  of  the  body,  the  lamp 
is  so  adjusted  that  a  fragment  of  cacao  butter  and  wax,  placed  close 
to  the  preparation,  is  melted. 

Israel's  Warming  Apparatus. — It  is  obvious  that  in  employing 
very  high  powers  a  difficulty  will  be  presented  by  the  warm  stages 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


125 


commonly   used    for   accurate   observations,    such    as    Schafer's   or 
Strieker's,  owing  to  their  interference  with  the  illumination.     To 


FIG.  53.— SECTION  OF  ISRAEL'S  WARMING  APPARATUS  AND  DROP-CULTURE 

SLIDE, 


overcome   this  an  apparatus   has  been  constructed   by  which    the 
slide  is  warmed  from  above  (Figs.  52,  53). 

The  drop-culture  slides  are  provided  with  a  shallow  groove,  *1  mm. 
deep  and  1  mm.  broad,  cut  round  the  concavity.  Into  this  the 
cover-glass  fits,  so  that  its  upper  surface  is  level  with  that  of 


FIG.  54. — ISRAEL'S  WARMING  APPARATUS. 

the  slide.     The  heating  apparatus  consists  of  a  flat  disk-shaped  box 
with  a  central  conical  aperture. 

The  entrance  and  exit  pipes  are  fixed  on  at  a  right  angle  to  the 


126  BACTERIOLOGY. 

side  (Fig.  54).  The  former,  z,  is  of  metal,  ancLthe  latter,  a,  of  glass 
fitted  with  a  thermometer,  the  bulb  of  which,  k,  is  contained  within 
the  box.  A  partition,  s,  keeps  up  a  current  between  the  openings 


FIG.  55. — GAS  CHAMBER  IN  USE  WITH  APPARATUS  FOR  GENERATING 
CARBONIC  ACID. 

of  the  pipes,   which  are  supported   on  a  stand   and  connected  by 
tubing  with  the  hot-water  supply. 

A  mixture  of  paraffine  and  vaseline  is  recommended  for  indicating 
the  temperature  of  the  chamber,  and  experience  has  shown  that  if 
a  temperature  of  37°  C.  is  required  the  temperature  of  the  water 
in  the  box  must  range  between  42°  and  47°  C. 


FIG.  56.— GAS  CHAMBER. 


Gas  Chambers. — To  investigate  the  action  of  gases  or  vapours 
upon  micro-organisms,  a  modification  of  the  moist  cell  may  be 
employed. 

A  piece  of  glass  tubing  is  first  fixed  to  the  slide  by  means  of 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


127 


sealing-wax,  and  the  ring  of  putty  is  so  placed  as  to  include  one  end 
of  it,  leaving  a  small  interval  at  the  side,  or  a  little  notch  is  made 
in  the  putty  opposite,  so  as  to  afford  an  exit  for  the  gas  or  vapour. 


FIG.  57.— MOIST  CELL  ADAPTED  FOR  TRANSMISSION  OF  ELECTRICITY. 

Application  of  Electricity. — To  study  the  effect  of  electricity 
we  may  prepare  a  drop-culture  in  the  moist  cell.  The  cover- 
glass  to  be  used  is  provided  with  two  strips  of  tinfoil,  which  are 


FIG.  58.— APPARATUS  ARRANGED  FOR  TRANSMITTING  ELECTRICITY. 

isolated  from  the  brass  of  the  microscope,  and  so  arranged  that  a 
current  of  electricity  may  be  passed  through  them. 

A  much  simpler  plan,  which  may  also  be  employed,  is  to  take 
an  ordinary  glass  slide  and  coat  the  surface  with  gold-size.     The 


128 


BACTERIOLOGY. 


slide  is  then  pressed  firmly  down  on  gold-leaf  or  tinfoil  and  allowed 
to  dry.  When  dry,  the  metal  is  scraped  away,  leaving  two  triangles 
with  a  small  interval  between  them. 


FIG.  59. — SLIDE  WITH  GOLD-LEAF  ELECTRODES. 

The  liquid  containing  the  micro-organisms  is  placed  between  the 
electrodes,  covered  with  a  cover-glass,  and  then  subjected  to  the 
electric  current. 

(G)  METHODS  OF  EMPLOYING  AND  STORING  LIQUID  MEDIA. 

Cultivations  in  liquid  media  can  be  carried  on  in  test-tubes,  but 
it  is  more  satisfactory  to  employ  special  forms  of  flasks,  bulbs,  and 
U  tubes,  such  as  those  employed  by  Pasteur  and  his  school,  and  by 
Lister,  Sternberg,  and  Aitken. 

Lister's  flasks. — These  flasks  were  especially  introduced  by  Lister 
as  a  means  of  storing  liquid  nutrient  media. 
They  are  so  constructed  that  after  removal 
of  a  portion  of  the  contents,  on  restoring 
the  vessel  to  the  vertical  position,  a  drop  of 
liquid  always  remains  in  the  extremity  of 
the  nozzle,  which  prevents  regurgitation  of 
unfiltered  air, 

Sternberg' s  Bulbs. — The  method  of  intro- 
ducing liquid  into  the.  bulb  employed  by 
Sternberg,  and  of  sterilising  and  inoculating 
it,  is  as  follows  :  The  bulb  is  heated  slightly 
over  the  flame,  and  the  extremity  of  the  neck,  after  the  sealed  point 
has  been  broken  off,  is  plunged  beneath  the  surface  of  the  liquid. 
As  the  air  cools  the  liquid  is  drawn  into  the  bulb, 

usually  filling  it  to  about  one-third  of  its  capacity.      C^. .— - — 

The  neck  of  the  flask  is  again  sealed  up,  and  the 

liquid  which  has  been  introduced  is  sterilised  by     FlG'  G|--STERX- 

BERG  s  BULB. 
repeatedly   boiling  the  flasks  in   the  water- bath. 

They  should  then  be  placed  in  the  incubator  for  two  or  three  days  ; 


FIG.  60.— LISTER'S 
FLASK. 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


129 


Fi 


62.— AITKEN'S 
TUBE. 


i\nd  if  the  contents  remain  transparent  and  free  from  film,  they 
may  be  set  aside  as  stock-bulbs,  to  be  used  when  required. 

To  inoculate  the  liquid  in  the  bulb  the  end  of  the  neck  is  heated 
to  sterilise  the  exterior,  the  bulb  is  gently  warmed,  and  the  extremity 
of  the  neck  nipped  off  with  a  pair  of  sterilised  forceps.  The  open 
extremity  is  plunged  into  the  liquid  containing  the  micro-organisms, 
and  a  minute  quantity  enters  the  tube  and  mingles  with  the  fluid  in 
the  bulb  without  fear  of  contamination  by  atmospheric  germs.  The 
extremity  of  the  neck  is  once  more  sealed  up  in  the  flame  of  a 
Bunsen  burner. 

Aitkeris  Tubes. — These  tubes  are  plugged  and  sterilised,  and  the 
nutrient  medium  introduced  as  into  ordinary  test-tubes.  Instead 
of  withdrawing  the  cotton-wool  plug,  they  are 
inoculated  through  a  lateral  arm.  The  sealed 
extremity  of  the  arm  is  nipped  off  with 
sterilised  forceps,  and  the  inoculating  needle  is 
carefully  introduced  through  the  opening  thus 
made.  It  is  directed  along  the  arm  until  it 
touches  the  opposite  side  of  the  test-tube,  where 
it  deposits  the  material  with  which  it  was  charged. 
The  needle  is  withdrawn,  and  the  end  of  the 
lateral  arm  again  sealed  up  in  the  flame ;  the  test-tube  is  then 
tilted  until  the  liquid  touches  the  deposited  material ;  on  restoring 
the  tube  to  the  vertical,  the  material  is  washed  down  with  the 
nutrient  liquid. 

Miguel's  Bulbs. — The  tube  a 
boule  of  Miquel  is  also  a  very 
useful  form.  It  consists  of  a 
bulb  of  50  cc.  capacity,  blown 
in  the  middle  of  a  glass  tube. 
The  part  of  the  tube  above  the 
bulb  is  contracted  in  two  places, 
and  can  either  be  left  quite 
straight  or  made  to  curve 
slightly.  Between  the  contrac- 
tions the  tube  is  plugged  with 
asbestos.  The  portion  of  the 
tube  below  the  bulb  is  $  shaped, 
and  drawn  out  at  its  extremity 

into  a  fine '  point.  The  bulb  is  charged  with  nutrient  liquid  and 
inoculated  by  aspiration,  and  the  point  of  the  $  tube  sealed  in  the 
flame  of  a  Bunsen  burner. 

9 


FIG.  63.— MIQUEL'S  BULB. 


130 


BACTERIOLOGY. 


FIG.  64. — PASTEUR'S  FLASK. 


Pasteur's  Apparatus. — Special  forms  of  tubes,  bulbs,  and  pipettes 
are  employed  by  the  school  of  Pasteur.     The  tubes  are  provided 

with  lateral  or  with 
curved  arms  drawn 
out  to  a  fine  point, 
and  with  slender 
necks  plugged  with 
cotton-wool.  A 
double  form  (Fig.  65) 
shaped  like  a  tuning- 
fork,  each  limb  with 
a  bent  arm,  is  con- 
venient for  storing 
sterilised  broth.  The 

sealed  end  of  an  arm  is  nipped  off  with  sterilised  forceps,  the  sterile 
broth  aspirated  into  each  limb,  and  the  arm  again  sealed  in  the 
flame ;  a  series  of  such  tubes  can  be 
arranged  upon  a  rack  on  the  working 
table. 

Bulbs  with  a  vertical  neck  drawn  out 
to  a  fine  point,  others  with  a  neck  bent 
at  an  obtuse  angle,  plugged  with  cotton- 
wool, and  a  lateral  curved  arm  drawn 
out  to  a  fine  point,  are  also  employed. 
For  a  description  of  these  various  vessels 
and  their  special  advantages,  the  works 

of   Pasteur   and    Duclaux    must   be  con- 

, .    ,  FIG.  65.— PASTEUR'S  DOUBLE 

suited.  TUM, 


(H)  CULTIVATION  OF  ANAEROBIC  BACTERIA. 

To  cultivate  anaerobic  organisms  the  same  media  are  employed 
as  for  aerobic  organisms,  but  the  methods  must  be  modified,  or  special 
apparatus  used,  so  that  the  oxygen  in  the  air  may  be  excluded. 

In  the  preparation  of  plate-cultivations,  before  the  film  of  gelatine 
has  completely  hardened  it  is  covered  with  a  sheet  of  mica,  arid  the 
edges  are  sealed  with  melted  paraffine.  By  this  process  the  air  is  not 
completely  excluded,  so  that  only  those  organisms  which  are  not 
strictly  anaerobic  can  be  grown  by  this  method.  Liborius  recom- 
mends boiling  a  considerable  volume  of  gelatine  in  a  tube,  cooling  it,. 
and  after  thoroughly  distributing  the  organisms  in  the  still  liquid  jelly, 
rapidly  solidifying  it  by  placing  the  tube  in  iced  water.  By  this 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


131 


process  very  little  air  re-enters  the  jelly,  and  colonies  of  even  strictly 
anaerobic  bacteria  will  develop  in  the  lower  part  of  the  tube.  The 
drawback  is  the  difficulty  encountered  in  examining  the  colonies, 
and  in  preparing  sub-cultures.  For  this  purpose  the  tube 
must  be  broken,  or  carefully  warmed  until  the  jelly  can  be 
shaken  out. 

Esmarch  first  prepares  a  roll  culture,  and  when  the  gelatine  film 
has  set,  the  tube  is  completely  filled  with  liquefied  gelatine  which  has 
been  cooled  down  almost  to  the  temperature  at  which  it  solidifies.  The 
same  difficulty  arises  as 
in  the  previous  method,  in 
the  examination  of  the 
colonies. 

Buchner  places  the 
culture  tube  inside  a  much 
larger  tube  containing  a 
small  quantity  of  pyro- 
gallic acid  and  closed  with 
a  gutta-percha  cap.  The 
pyrogallic  acid  absorbs  the 
oxygen,  but  the  method 
is  not  altogether  success- 
ful. 

The  most  satisfactory 
plan  is  to  exhaust  the  air 
with  an  air  pump,  or  to 
substitute  an  atmosphere 
of  hydrogen  which  does 
not  affect  the  growth  of 
the  bacteria. 

Various  forms  of  flasks 
and  tubes  for  cultivating 
bacteria  have  been  devised, 
which  can  be  easily  con- 
nected with  an  exhausting 


66>_FRA-NKEL,S  ANAEROBIC   TUBE-CULTURE' 
Glass   tube   through    which    hydrogen    is 
passed;     b,     exit      tube;     c,     india-rubber 
stopper     coated     externally     with    paraffin 
(FRANKLAND). 


apparatus,      and     readily 

sealed  by  the  flame  of  the  blowpipe  when  the  air  has  been  removed. 

If  hydrogen  is  employed  the  most  convenient  plan  is  to  use 
a  Kipp's  apparatus,  from  which  the  hydrogen  is  passed  through 
two  bottles,  one  containing  a  solution  of  lead,  to  remove  any 
sulphuretted  hydrogen,  and  the  other  pyrogallic  acid,  to  intercept 
any  oxygen. 


132 


BACTERIOLOGY. 


In  the  method  recommended  by  Frankel  a  tube  of  gelatine  is 
liquefied,  and  inoculated.  A  gutta-percha  stopper  is  substituted 
for  the  cotton-wool  plug  (Fig.  66).  It  is  perforated  by  two  holes, 
through  which  two  tubes  pass  which  are  bent  at  a  right  angle. 
One  tube  only  just  passes  through  the  stopper,  the  other  reaches 
down  to  the  bottom  of  the  test-tube.  The 
horizontal  part  of  each  tube  has  a  narrow 
neck.  The  long  tube  has  a  plug  of  steril- 
ised cotton-wool,  and  is  connected,  with 
a  short  piece  of  india-rubber  tubing  by 
which  it  can  be  connected  with  Kipp's 
apparatus.  The  hydrogen  drives  the  air 
out  of  the  liquefied  jelly  and  out  of  the 
test-tube,  and  after  about  half  an  hour 
the  horizontal  tubes  are  sealed  up,  and 
the  test-tube  is  made  into  a  roll  culture. 

Liborius  employs  a  tube  with  a  narrow 
neck  and  a  lateral  arm  (Fig.  67).  The 
tube  is  filled  up  to  the  height  of  the  arm 
with  either  nutrient  agar  or  a  mixture  of 
nutrient  agar  with  2  per  cent,  of  grape- 
sugar.  The  liquefied  jelly  is  inoculated 
in  the  usual  way,  and  hydrogen  passed 
through  the  lateral  arm.  When  the  air 
has  been  completely  driven  out,  the  tube 
is  sealed  up. 

To  cultivate  anaerobic  organisms  in 
broth,  such  as  the  tetanus  bacillus,  a  flask 
is  inoculated  with  the  bacillus,  and  a 
stream  of  hydrogen  is  passed  through  the 
broth  by  means  of  a  tube  passing  down  to  the  bottom  of  the 
flask.  The  air  in  the  flask  escapes  by  a  lateral  arm  which  is  bent 
down  at  a  right  angle,  and  immersed  in  a  capsule  of  mercury. 
When  the  air  has  been  completely  expelled  the  entrance  tube  is 
hermetically  sealed,  and  the  mercury  in  the  capsule  prevents  any 
air  from  re-entering  the  flask  by  the  lateral  arm  (Fig.  68). 


FIG.  67.— ANAEROBIC  CUL- 
TURE-TUBE (LlBORIUS). 


METHOD  OF  FIXING  CULTURES. 

The  colonies  in  plate -cultivations  and  the  growths  of  bacteria 
in  test-tubes  may  be  stopped  at  any  stage  of  their  growth,  and 
permanently  fixed  by  exposing  the  culture  to  the  fumes  of  formic 


NUTRIENT   MEDIA   AND   METHODS   OF   CULTIVATION. 


133 


aldehyde.     The  test-tubes,  dishes,  or 'capsules  are  placed  in  a  cylin- 
drical glass  vessel  containing  a  pledget  of  cotton-wool  moistened  with 


FIG.    68. — APPAKATUS   FOR    ANAEROBIC    CULTURES. 

(ROSCOE  AND  LUNT.) 

formic  aldehyde.  The  vessel  is  fitted  with  a  ground  glass  stopper 
and  set  aside.  The  growth  almost  immediately  ceases.  Any 
liquefied  gelatine  is  hardened,  so  that  the  exact  appearances  of 
cultures  are  obtained  in  a  permanent  form. 


CHAPTER  X. 

EXPERIMENTS   UPON   THE   LIVING   ANIMAL. 

To  carry  out  the  last  of  Koch's  postulates,  and  so  complete  the 
chain  of  evidence  in  favour  of  the  causal  relation  of  micro-organisms 
to  disease,  and  to  study  the  mode  of  action  of  a  pathogenic  bacterium, 
it  is  necessary  to  introduce  into  a  living  animal  a  pure  cultivation 
of  the  micro-organism  or  its  chemical  products.  For  this  purpose 
various  animals  are  employed,  such  as  mice,  rabbits,  guinea-pigs, 
pigeons,  and  fowls. 

Inhalation. — The  animals  may  be  made  to  inhale  an  atmosphere 
impregnated  with  micro-organisms  by  means  of  a  spray.  In  this 
way  Priedlander  succeeded  in  administering  the  bacteria  of  pneu- 
monia to  mice ;  and  the  production  of  tuberculosis  by  experimental 
inhalation  has  thrown  light  upon  the  clinical  records  of  cases 
reported  as  instances  o£  the  infectiousness  of  phthisis. 

Ingestion. — A  sheep  fed  upon  potatoes  which  have  been  the 
medium  for  the  cultivation  of  the  anthrax  bacillus  dies  in  a  few 
days.  Rabbits  fed  on  cabbage  sprinkled  with  a  culture  of  the 
bacillus  of  fowl  cholera,  rapidly  succumb  to  the  disease.  Animals 
fed  upon  the  nodules  of  bovine  tuberculosis  or  upon  tubercular 
flesh  and  milk  will  be  readily  infected. 

Milk,  or  bread  soaked  in  milk,  is  a  very  convenient  medium, 
and  from  a  public  health  point  of  view,  a  most  instructive  way  of 
administering  and  testing  the  effect  of  pathogenic  bacteria. 

Vaccination  and  Subcutaneous  Inoculation. — Vaccination  may  be 
performed  by  making  a  few  superficial  scratches  and  inoculating  the 
wound  with  a  sterilised  platinum  needle  charged  with  the  micro- 
organisms. Another  simple  method  is  to  take  a  sterilised  scalpel, 
infect  the  point  with  the  material  to  be  inoculated,  and  then  make 
a  minute  puncture  or  incision.  In  either  case  a  situation  should  be 
selected,  such  as  the  root  of  the  ear,  which  cannot  be  licked  by  the 
animal  after  the  operation. 

134 


EXPERIMENTS    UPON    THE    LIVING    ANIMAL. 


135 


Subcutaneous  inoculation  is  very  simple  and  effectual,  and  con- 
sequently the  method  most  frequently  employed.  The  animal 
selected — for  example,  a  guinea-pig — is  held  by  an  assistant,  who 
covers  it  with  a  towel,  leaving  only  the  hind  extremities  exposed. 
By  so  doing,  and  gently  laying  it  upon  its  back,  with  its  head  low, 
a  guinea-pig  passes  apparently  into  a  state  of  hypnotism,  and  the 


FIG.  69.— KOCH'S  SYRINGE. 

trivial  operation  can  be  performed  with  little  or  no  movement  on  the 
part  of  the  animal.  From  a  spot  on  the  inner  side  of  the  thigh  the 
hair  is  cut  close  with  a  small  pair  of  scissors  curved  on  the  flat,  and 
the  skin  must  be  thoroughly  purified  with  1  in  20  carbolic  acid.  A 
small  fold  of  skin  is  then  pinched  up  with  a  pair  of  sterilised  forceps, 
and  with  a  pair  of  sharp  sterilised  scissors,  or  with  a  tenotomy  knife, 
a  minute  incision  is  made.  A  sterilised  platinum  loop  is  charged 
with  the  material  to  be  inoculated,  and  the  loop  is  gently  inserted 
under  the  skin,  forming  a  small  pocket  in  the  subcutaneous  tissue. 
The  needle  is  then  withdrawn,  and  the  sides  of  the  wound  gently 
pressed  into  apposition  and  painted  over  with  collodion. 


FIG.  70.— SYRINGE  WITH  ASBESTOS  PLUG. 

In  inoculating  a  mouse  the  same  process  is  adopted,  with  the 
exception  that  the  root  of  the  tail  is  the  usual  site  of  the 
operation. 

In  some  cases  it  may  be  necessary  to  inoculate  cultures  diffused 
in  sterilised  salt  solution,  or  blood  or  lymph  containing  bacteria, 
or  a  culture  in  broth,  or  a  filtrate  containing  the  toxic  products, 


136  BACTERIOLOGY. 

and  then  a  hypodermic  syringe  may  be  required.  One  of  the  ordinary 
pattern  may  be  used,  but  it  is  very  much  better  to  employ  a  syringe 
which  has  been  especially  constructed  to  admit  of  thorough  dis- 
infection. Koch's  syringe  is  a  convenient  form,  the  liquid  being 
expressed  by  pressure  on  a  rubber  ball. 

The  author  has  generally  preferred  to  improvise  a  substitute  for 
the  hypodermic  syringe  which  can  be  quickly  made,  and  is  destroyed 
after  use,  so  that  there  can  be  no  possible  risk  of  accidentally 
infecting  other  animals.  A  short  length  of  ordinary  glass-tubing  is 
sterilised,  and  plugged  at  one  end  with  sterilised  cotton- wool ;  about 
three  inches  from  the  plug  a  bulb  is  blown  about  the  size  of  a 
marble,  and  two  inches  below  this  the  glass  is  drawn  out  into  a  long 
capillary  tube.  A  sufficient  quantity  of  the  liquid  to  be  injected  rises 
up  into  the  tube  by  capillary  attraction,  or  can  be  drawn  up  by 
means  of  an  india-rubber  ball,  until  the  bulb  is  full.  The  point  of 
the  capillary  tube  is  inserted  through  the  opening  in  the  skin,  and 
gently  pushed  into  the  subcutaneous  tissue,  and  then  withdrawn  for 
a  short  distance.  By  pressure  on  the  bulb  the  contents  of  the  tube 
are  injected.  In  dealing  with  chemical  products  there  is  no  risk  in 
applying  the  lips  and  blowing  out  the  contents  of  the  tube,  or  indeed 
of  filling  it  by  suction,  for  if  too  much  force  were  applied  the  liquid 
which  might  enter  the  mouth  would  be  stopped  by  the  cotton-wool 
plug. 

A  number  of  these  capillary  tubes  can  be  placed  in  a  small  case, 
and  when  it  is  necessary  to  go  to  a  distance  to  investigate  an  outbreak, 
they  will  be  found  most  convenient  to  bring  back  lymph  or  blood  to 
the  laboratory  for  further  study. 

Sternberg  takes  a  piece  of  glass-tubing,  blows  a  bulb  at  one  endr 
and  draws  out  the  other  end  into  a  thin  tube.  By  heating  the  bulb 
and  then  dipping  the  tube  into  the  liquid  to  be  inoculated  the  latter 
rises  in  the  tube  as  the  bulb  cools.  After  inserting  the  point  of 
the  tube  subcutaneously  the  bulb  is  again  heated,  and  the  liquid  is 
forced  out  into  the  tissues. 

Intravenous  Injection. — A  cultivation  of  micro-organisms  may  be 
mixed  with  sterilised  water,  and  then  injected  with  a  syringe  directly 
into  the  circulation.  This  may  be  performed  without  much  difficulty 
by  injecting,  with  a  hypodermic  syringe,  the  large  vein  at  the  base 
of  the  ear  in  rabbits,  or  the  jugular  vein  in  large  animals. 

Special  Operations. — In  many  cases  it  is  absolutely  necessary  to 
perform  an  operation  of  greater  severity.  After  the  administration 
of  an  anaesthetic,  infective  material  may  be  inserted,  or  injected,  into 
the  peritoneal  cavity,  or  injected  into  the  duodenum  in  the  manner 


EXPERIMENTS   UPON   THE   LIVING   ANIMAL.  137 

employed  in  the  case  of  Koch's  comma  bacilli  by  Nicati  and  Bietsch. 
In  such  cases  antiseptic  precautions  must  be  rigidly  followed,  and 
use  made  of  iodoform  and  other  antiseptic  dressings.  The  disinfec- 
tion of  the  skin  of  the  animal,  of  the  instruments  employed,  and  of 
the  hands  of  the  operator,  are  details  essential  to  secure  success. 

To  inoculate  tubercular  matter,  sputum  may  be  rubbed  up  with 
distilled  water,  and  some  of  the  mixture  injected  into  a  tracheal 
fistula  ;  or  the  first  steps  of  the  operation  of  iridectomy  may  be 
performed  and  tubercular  material  inserted  into  the  anterior  chamber 
of  the  eye,  but  this  method  is  only  justifiable  when  it  is  absolutely 
necessary  for  the  results  and  changes  to  be  observed  from  day 
to  day. 

To  inoculate  rabbits  or  other  animals  with  the  virus  of  rabies, 
the  skull  is  trephined,  and  an  emulsion  prepared  from  the  spinal  cord 
of  a  rabid  animal  is  injected  beneath  the  dura  mater. 

Before  every  inoculation  the  instruments  must  be  sterilised  in  a 
hot-air  steriliser  or  by  immersion  in  boiling  water  in  a  flat  dish  or 
enamel  tray  heated  by  a  spirit-lamp,  and  after  each  operation  all 
instruments  should  be  placed  in  carbolic  acid  (1  in  20)  or  in  boiling 
water,  wiped  dry,  and  again  sterilised  in  the  hot-air  steriliser,  before 
they  are  put  away.  If  these  precautions  are  not  observed,  instances 
of  accidental  infection  are  sure  to  occur. 

After  the  inoculation  is  completed  a  careful  record  must  be  made 
of  the  date  and  details  of  the  experiment.  The  form  in  which  the 
virus  was  used,  the  quantity  employed,  and  the  seat  of  inoculation, 
must  be  taken  into  account.  The  animals  must  be  kept  under  close 
observation,  the  temperature  taken,  and  any  signs  of  illness,  such  as 
ceasing  to  feed,  difficulty  in  breathing,  staring  coat,  and  any  local 
signs,  such  as  the  development  of  a  tumour  or  an  enlargement  of  the 
lymphatic  glands,  must  be  carefully  noted. 

It  is  perhaps  hardly  necessary  to  add  that  in  this  country  no 
experiments  of  any  kind  may  be  performed  on  living  animals  without 
a  license. 

METHOD  OF  DISSECTION  AND  EXAMINATION. 

All  animals  that  die  after  an  experimental  inoculation  should 
be  examined  immediately  after  death.  Every  precaution  must  be 
taken  in  conducting  the  dissection,  to  exclude  extraneous  micro- 
organisms, and  all  instruments  employed  must  have  been  sterilised 
in  the  hot-air  steriliser,  or  by  immersion  in  boiling  water.  If  a  mouse, 
for  example,  has  died  after  inoculation  with  anthrax,  it  should  be  at 


138  BACTERIOLOGY. 

once  pinned  out  by  its  feet  on  a  slab  of  wood  or  in  a  gutta-percha 
tray,  and  bathed  with  1  in  40  carbolic.  In  the  same  way,  before 
examining  a  dead  rabbit,  a  stream  of  carbolic  should  be  directed, 
over  it  to  lay  the  fur,  which  otherwise  interferes  with  the  dissection. 
The  hair  should  be  cat  away  with  sterilised  scissors  from  the  seat 
of  inoculation,  which  is  the  first  part  to  be  examined,  and  any 
suppuration,  haemorrhage,  redema,  or  other  pathological  change  should 
be  carefully  noted.  From  any  pus  or  exudation  that  may  be 
present,  material  for  inoculations  should  at  once  be  taken,  and 
cover-glass-preparations  made  for  microscopical  examination. 

To  examine  the  internal  organs  and  to  make  inoculations  from 
the  blood  of  the  heart  or  spleen,  the  skin  is  cut  through  from  below 
upwards  in  the  median  line  of  the  abdominal  and  thoracic  regions. 
The  abdominal  cavity  is  then  opened,  and  the  walls  pinned  back 
on  either  side  of  the  animal.  Any  abnormal  appearances  in  the 
peritoneum  should  be  noted,  and  the  state  of  the  spleen  should  be 
carefully  examined  by  turning  the  intestines  aside.  After  noting 
its  appearances,  it  should  be  removed  with  sterilised  forceps  and 
scissors,  and  deposited  upon  a  sterilised  glass  slide,  and  incised  with 
sterilised  scissors.  The  cut  surface  is  then  touched  with  the  point 
of  a  sterilised  inoculating  needle,  and  cultures  are  made  in  test-tubes 
of  nutrient  gelatine  and  agar-agar,  and  also  on  potato,  and  in  broth 
in  the  form  of  drop- cultivations.  Precisely  the  same  care  must  be 
taken  in  examining  lymphatic  glands,  tubercles,  or  pathological 
nodules  ;  any  chance  putrefactive  micro-organisms  on  the  surface 
should  be  destroyed  by  carbolic  acid  or  the  actual  cautery;  an 
incision  is  then  made,  and  a  minute  fragment  snipped  out  of  the 
centre  of  the  nodule,  which  can  be  inoculated  in  the  living  animal  or 
transferred  to  a  cultivating  medium. 

The  examination  of  the  thorax  is  made  by  cutting  through  the 
ribs  on  either  side  of  the  sternum  with  sterilised  scissors,  and 
turning  the  sternum  up  where  it  will  be  out  of  the  way.  The 
pericardium  is  then  opened,  and  the  right  auricle  or  ventricle  pierced 
with  the  point  of  a  sterilised  scalpel,  and  inoculations  and  cover- 
glass-preparations  are  made  from  the  blood  which  escapes. 

The  lungs  also  require  to  be  especially  studied.  They  should  be 
incised  with  a  sterilised  scalpel,  and  inoculations  and  cover-glass- 
preparations  made  from  the  cut  surface.  It  may  be  necessary  to 
embed  a  piece  of  lung  or  fragment  of  spleen,  so  that  it  shall  be  free 
from  air.  This  may  be  done  by  isolating  a  fragment  with  the 
precautions  just  described,  and  depositing  it  upon  the  surface  of  a 
test-tube  of  nutrient  agar-agar.  The  contents  of  another  tube, 


EXPERIMENTS   UPON   THE   LIVING   ANIMAL.  139 

which  have  been  liquefied,  and  allowed  to  cool  almost  to  the  point 
of  gelatinisation,  must  then  be  poured  over  it.  From  a  potato  a 
little  cube  must  be  cut,  the  tissue  deposited  in  the  trough  thus 
formed,  and  the  cube  replaced,  or  cultures  may  be  prepared  by  any 
of  the  methods  which  have  been  described  for  dealing  with  anaerobic 
bacteria.  Blood  may  also  be  taken  directly  from  a  vein  by  laying  it 
bare  by  dissection,  making  a  small  opening  with  sterilised  scissors, 
and  inserting  a  looped  platinum  needle,  the  needle  of  a  hypodermic 
syringe,  a  capillary  tube,  or  the  extremity  of  the  capillary  neck  of 
a  Sternberg's  bulb.  If  the  cultivation,  in  spite  of  these  precautions, 
is  contaminated,  or  if  there  was  more  than  one  organism  present 
in  the  blood  or  tissues  under  examination,  it  will  be  necessary  to 
separate  the  different  kinds  by  plate -cultivation. 

Having  completed  the  dissection,  the  organs  of  such  a  small 
animal  as  a  mouse  may  be  removed  en  masse,  and  transferred  to 
absolute  alcohol  for  subsequent  examination.  In  other  cases  it  may 
be  only  necessary  to  reserve  portions  of  each  organ.  In  experimenting 
with  a  virulent  micro-organism  like  anthrax,  any  remaining  part  of 
the  animal  should  be  cremated,  and  the  hands  and  all  instruments 
should  be  thoroughly  disinfected. 

Isolation  of  Micro-organisms  during  Life. — Micro-organisms  in 
the  living  subject  may  be  isolated  from  the  pus  of  abscesses,  or 
other  discharges,  and  from  the  blood  and  tissues.  Abscesses  should 
be  opened,  and  other  operations  performed,  when  practicable,  with 
Listerian  precautions,  and  a  drop  of  the  discharge  taken  up  with 
Ji  looped  needle  or  capillary  pipette,  as  already  explained. 

To  make  a  cultivation  from  the  blood  of  a  living  person,  the  tip 
of  a  finger  must  be  well  washed  with  soap  and  water  and  sponged 
with  1  in  20  carbolic.  Venous  congestion  is  produced  by  applying 
an  elastic  band  or  ligature  to  the  finger,  which  is  pricked  with  a 
sterilised  sewing  needle.  From  the  drop  of  blood  which  exudes  the 
necessary  inoculations  and  examinations  can  be  made.  Another 
way  of  extracting  blood  from  the  living  patient  is  to  apply  a  leech. 
This  method  has  been  found  of  considerable  value  in  experimenting 
upon  the  blood  of  patients  suffering  from  malaria,  and  may  be 
useful  in  other  diseases,  if  the  blood  is  required  for  further 
examination,  or  in  quantity. 


CHAPTER   XI. 

EXAMINATION   OF  AIR,    SOIL,    AND   WATER. 

AlR. 

THE  air,  as  is  well  known,  contains  in  suspension,  mineral,  animaly 
and  vegetable  substances.  The  mineral  world  is  represented 
by  such  substances  as  silica,  silicate  of  aluminium,  carbonate  and 
phospate  of  calcium,  which  may  be  raised  from  the  soil  by  the 
wind,  and  particles  of  carbon,  etc.,  which  gain  access  from  acci- 
dental sources.  Belonging  to  the  animal  kingdom  we  find  the 
debris  of  perished  creatures,  as  well  as,  sometimes,  living  animals. 
The  vegetable  world  supplies  micrococci,  bacilli,  and  other  forms 
of  the  great  family  of  bacteria,  spores  of  other  fungi,  pollen  seeds, 
parts  of  flowers,  and  so  forth.  The  air  of  hospitals  and  sick  rooms 
has  been  found  to  be  especially  rich  in  vegetable  forms;  fungi 
and  spores  have  been  stated  to  be  present  in  particularly  large 
numbers  in  cholera  wards;  spores  of  tricophyton  have  been  dis- 
covered in  the  air  of  hospitals  for  diseases  of  the  skin,  and  of  achorioii 
in  wards  with  cases  of  favus.  The  tubercle  bacillus  is  said  to  have 
been  detected  in  the  breath  of  patients  suffering  from  phthisis. 

These  points  indicate  that,  in  addition  to  the  interest  for  the 
micro- biologist,  considerable  importance,  from  a  hygienic  point  of 
view,  must  be  attached  to  the  systematic  examination  of  the  air. 
A  knowledge  of  the  microbes  which  are  found  jn  the  air  of  marshy 
and  other  unhealthy  districts,  and  in  the  air  of  towns,  dwellings, 
hospitals,  workshops,  factories,  and  mines,  will  be  of  practical  value. 

Miquel,  who  has  particularly  studied  the  bacteria  in  the  air, 
has  found  that  their  number  varies  considerably.  The  average 
number  per  cubic  metre  of  air  for  the  autumn  quarter  at  Mont- 
souris  is  given  as  142,  winter  quarter  49,  spring  quarter  85,  and 
summer  quarter  105.  In  air  collected  2,000  to  4,000  metres  above 
the  sea-level,  not  a  single  bacterium  or  fungus  spore  was  found, 
while  in  10  cubic  metres  of  air  from  the  Rue  de  Rivoli  (Paris)  the 
number  was  computed  at  55,000. 

140 


EXAMINATION   OF   AIR,    SOIL,   AND   WATER.  141 

The  simplest  method  for  examining  the  organisms  in  air  consists 
in  exposing  plates  of  glass  or  microscopic  slides  coated  with 
glycerine,  or  with  a  mixture  of  glycerine  and  grape  sugar,  which  is 
stable,  colourless,  and  transparent.  Nutrient  gelatine  spread  out  on 
glass  plates  may  be  exposed  to  the  air  for  a  certain  time,  and  then 
put  aside  in  damp  chambers  for  the  colonies  to  develop.  Sterilised 
potatoes,  prepared  in  the  usual  way,  may  be  similarly  exposed.  In 
both  the  last- mentioned  methods  separate  colonies  develop,  which 
may  be  isolated,  and  pure  cultivations  carried  on  in  various  other 
nutrient  media.  Nutrient  gelatine  has  also  been  employed  in  the 
special  methods  of  Koch  and  Hesse. 

Koch's  Apparatus. — This  consists  of  a  glass  jar,  about  six  inches 
high,  the  neck  of  which  is  plugged  with  cotton-wool.  In  the 
interior  is  a  shallow  glass  capsule,  which  can  be  removed  by  means 
of  a  brass  lifter.  The  whole  is  sterilised  by  exposure  to  150°  C. 
for  an  hour  in  the  hot-air  steriliser.  The  nutrient  gelatine  in  a 
stock-tube  is  liquefied,  and  the  contents  emptied  into  the  glass 
•capsule.  The  jar  is  exposed  to  the  air  to  be  examined  for  a  definite 
time,  the  cotton -wool  plug  replaced,  and  the  apparatus  set  aside  for 
the  colonies  to  develop. 

Hesse's  Apparatus. — The  advantage  of  this  apparatus  is  that  it 
enables  the  experimenter  to  examine  a  known  volume  of  air.  A 
glass  cylinder,  70  cm.  long  and  3*5  cm.  in  diameter,  is  closed  at  one 
end  by  an  india-rubber  cap,  perforated  in  the  centre.  Over  this  fits 
another  cap,  which  is  not  perforated.  The  opposite  end  of  the 
cylinder  is  closed  with  a  caoutchouc  stopper,  perforated  to  admit 
a  glass  tube  plugged  with  cotton-wool.  The  tube  can  be  connected 
by  means  of  india-rubber  tubing  with  an  aspirating  apparatus, 
which  consists  of  a  couple  of  litre-flasks,  suspended  by  hooks  from 
the  tripod-stand  which  supports  the  whole  apparatus.  The  cylinder, 
caps,  and  plug  are  washed  with  solution  of  carbolic  acid,  and 
then  with  alcohol.  After  being  thus  cleansed,  50  cc.  of  nutrient 
gelatine  are  introduced,  and  the  whole  sterilised  by  steaming  for 
half  an  hour  for  three  successive  days.  After  the  final  sterilisation, 
the  cylinder  is  rotated  on  its  long  axis,  so  that  the  nutrient  medium 
solidifies  in  the  form  of  a  coating  over  the  whole  of  the  interior. 
When  required  for  use,  the  cotton-wool  plug  is  removed  from  the 
small  glass  tube,  and  the  latter  connected  with  the  upper  flask  by 
means  of  the  india-rubber  tubing. 

The  apparatus  is  placed  in  the  air  which  is  to  be  examined,  the 
outer  india-rubber  cap  removed  from  the  glass  cylinder,  and  the 
upper  flask  tilted  until  the  water  begins  to  flow  into  the  lower  one. 


142 


BACTERIOLOGY. 


The  emptying  continues  by  siphon  action,  and  air  is  drawn  in  along 
the  cylinder  to  replace  the  water.  When  the  upper  flask  is  empty, 
the  position  of  the  two  is  reversed,  and  the  flow  again  started. 
When  a  sufficient  volume  has  been  drawn  through  the  cylinder,  the 
outer  cap  and  the  cotton-wool  plug  are  replaced,  and  it  is  set  aside 
for  the  colonies  to  develop.  As  an  example,  twenty-five  litres  of  air 
from  an  open  square  in  Berlin  gave  rise  to  three  colonies  of  bacteria 
and  sixteen  moulds;  on  the  other  hand,  two  litres  from  a  school  - 


FIG.  71.— HESSE'S  APPARATUS.    • 

room  just  vacated  by  the  scholars  gave  thirty-seven  colonies  of 
bacteria  and  thirty-three  moulds. 

Porous  substances,  such  as  sand,  powdered  glass,  or  sugar,  may 
be  used  for  the  filtration  of  samples  of  air ;  and  an  apparatus  is 
employed  in  a  convenient  form  to  be  conveyed  to  the  laboratory  for 
the  subsequent  examination. 

Petri's  Apparatus  consists  of  a  glass-tube  9  cm.  long,  containing 
two  sand-filters  separated  from  each  other.  A  known  volume  of  air 
is  aspirated  through  the  tube.  The  bacteria  are  arrested  and  can 


EXAMINATION   OF   AIR,    SOIL,    AND   WATER. 


143 


be  examined  by  spreading  the  sand  out  in  a  dish  and  covering  it 
with  nutrient  gelatine ;  or  it  may  be  shaken  up  with  sterilised  water 
and  pi  ate -cultivations  prepared.  The  sand-filter  nearest  to  the 
aspirator  should  remain  free  from  bacteria. 

Sedywick  and  Tucker  employ  a  glass  cylinder  which  is  drawn  out 
at  one  end  into  a  narrow  tube  to  contain  sterilised  powdered  cane 
sugar.  Both  ends  of  the  apparatus  are  plugged  with  sterilised 
cotton-wool.  By  means  of  an  exhausting  apparatus  a  known  volume 
of  air  is  drawn  through  the  tube.  The  cotton-wool  plug  is  re- 


FIG.  72. — SEDGWICK  AND  TUCKER'S  TUBE. 

moved,  and  liquid  gelatine  is  introduced  into  the  cylinder,  the 
plug  is  replaced,  and  the  sugar  is  shaken  into  and  quickly  dissolves 
in  the  jelly.  The  cylinder  is  then  treated  in  the  same  way  as  a 
roll-culture,  and  set  aside  for  the  colonies  to  develop  (Fig.  72). 


Various  forms  of  "  aeroscopes  "  and 


"  aeroniscopes  "  have  from 


FIG.  73.— POUCHET'S  AEROSCOPE. 


time  to  time  been  employed.     Pouchet's  aeroscope  consists  of  a  small 
funnel,  drawn  out  to  a  point  below  which  is  a  glass  slip  coated  with 


144  BACTERIOLOGY. 

glycerine.  The  end  of  the  funnel  and  the  glass  slip  are  enclosed 
in  an  air-tight  chamber,  from  which  a  small  glass  tube  passes  out 
and  is  connected  by  india-rubber  tubing  with  an  aspirator  (Fig.  73). 
The  air  passing  down  the  funnel  strikes  upon  the  glycerine,  which 
arrests  any  solid  particles.  For  a  full  description  of  the  apparatus 
employed  by  Maddox,  Cunningham,  and  Miquel,  reference  should 
be  made  to  the  writings  of  these  authors,  and  particularly  to  the 
treatise  published  by  the  last-named. 

SOIL. 

Surface  soil  is  exceedingly  rich  in  bacteria.  Miquel  has  com- 
puted that  there  exists  in  a  gramme  of  soil  an  average  of  750,000 
germs  at  Montsouris,  1,300,000  in  the  Rue  de  Rennes,  and  2,100,000 
in  the  Rue  de  Monge.  As  agents  in  putrefaction  and  fermenta- 
tion they  play  a  very  important  rdle  in  the  economy  of  nature ; 
but  there  exist  in  addition,  bacteria  in  the  soil  which  are  patho- 
genic in  character.  Pasteur  has  succeeded  in  isolating  the  bacillus 
of  anthrax  from  the  earth.  Sheep,  sojourning  upon  a  plot  of 
ground  where  animals  with  anthrax  have  been  buried,  may  succumb 
to  the  disease.  Pasteur  considered  that  the  spores  were  conveyed 
by  worms  from  buried  carcasses  to  the  surface  soil.  The  bacilli 
of  malignant  redema  and  tetanus  are  also  present  in  soil.  Nicolaier 
produced  tetanus  in  mice  and  rabbits  by  inoculating  a  little  garden 
earth  under  the  skin. 

To  obtain  a  cultivation  of  the  microbes  in  soil  a  sample  of  the 
latter  must  be  first  dried  and  then  triturated.  It  may  then  be 
shaken  up  with  distilled  water,  and  from  this  a  drop  transferred  to 
sterilised  broth.  The  employment  of  solid  media  is,  however, 
much  more  satisfactory :  a  sample  of  earth  is  collected,  dried,  and 
triturated,  and  a  small  quantity  sprinkled  over  the  surface  of 
nutrient  gelatine  prepared  for  a  plate-cultivation.  In  another 
method  the  gelatine  is  liquefied  in  a  test-tube,  the  powder  added, 
and  distributed,  in  the  usual  way,  throughout  the  medium,  which 
is  then  poured  out  upon  a  glass  plate  or  made  into  a  roll-culture. 
In  the  same  way  the  dust  which  settles  from  the  air  in  houses  and 
hospitals,  or  food  substances  in  powder,  may  be  distributed  in 
nutrient  gelatine,  and  examined  both  for  aerobic  and  anaerobic 
bacteria.  The  different  kinds  which  develop,  must  be  thoroughly 
investigated  as  regards  their  morphological  and  biological  charac- 
ters, and  pathogenic  properties. 


EXAMINATION   OF  AIR,   SOIL,   AND   WATER.  145 

WATER. 

In  the  case  of  water,  as  in  that  of  air,  a  knowledge  of  the 
micro-organisms  which  may  be  present  is  not  only  of  interest 
to  the  bacteriologist,  but  of  the  greatest  importance  in  practical 
hygiene.  Common  putrefactive  bacteria  and  vibrios  may  not  be 
hurtful  in  themselves,  but  they  indicate  the  probability  of  the 
presence  of  organic  matter  in  which  there  may  be  danger.  The 
detection  of  Bacillus  coli  communis  may  be  taken  to  indicate  a 
probable  contamination  with  human  excreta. 

The  Microzyme  Test  which  was  introduced  for  the  detection 
of  putrefactive  bacteria,  consisted  in  adding  three  or  four  drops  of 
the  sample  of  water  to  1  or  2  cc.  of  Pasteur's  fluid,  the  nourishing 
fluid  having  been  previously  boiled  in  a  sterilised  test-tube.  If  the 
microzymes  or  their  germs  existed  in  the  water,  the  liquid  in  a  few 
days  became  turbid  from  the  presence  of  countless  bacteria.  This 
test  is  of  110  real  value,  for  it  does  little  more  than  indicate  that 
bacteria  are  present,  which  we  know  to  be  the  case  in  all  ordinary 
water,  and  even  in  ice.  On  the  other  hand,  the  bacteriological  test 
of  Koch  is  a  most  valuable  addition  to  the  usual  methods  of  water 
analysis.  It  enables  us  not  only  to  detect  the  presence  of  bacteria, 
but  to  ascertain  approximately  their  number,  and  to  study  very 
minutely  their  morphological  and  biological  characteristics.  The 
importance  of  a  thorough  acquaintance  with  the  life- history  of  the 
individual  micro-organisms  cannot  be  too  strongly  insisted  upon. 
For  example,  by  such  means  the  spirillum  of  Asiatic  cholera  can 
be  distinguished  from  most  other  comma-shaped  organisms,  and 
inasmuch  as  its  presence  may  be  an  indication  of  contamination 
with  choleraic  discharges,  such  water  should  be  condemned  for 
drinking  purposes,  even  though  we  are  not  yet  in  a  position  to 
affirm  that  the  microbe  is  the  cause  of  the  disease.  The  detection 
of  the  bacillus  of  typhoid  fever  or  of  the  Bacillus  coli  communis 
in  suspected  water  or  milk  would  be  evidence  of  considerable 
importance. 

Koch's  test,  in  short,  consists  in  making  plate-cultivations  of  a 
known  volume  of  water,  counting  the  colonies  which  develop f 
isolating  the  micro-organisms,  and  studying  the  characters  of  each 
individual  form. 

Collection  and  Transport  of  Water  Samples. — Sternberg's  bulbs, 
or  Erlenmeyer's  conical  flasks  of  about  100  cc.  capacity,  may  be 
employed  with  advantage  for  collecting  the  samples  of  water.  The 
latter  are  cleansed,  plugged,  and  sterilised  in  the  hot-air  steriliser. 

10 


146  BACTERIOLOG  Y . 

When  required  for  use,  the  plug  is  removed  and  held  between  the 
fingers,  which  must  not  touch  the  part  which  enters  the  neck  of 
the  flask.  About  30  cc.  of  the  water  to  be  examined  are  intro- 
duced into  the  flask,  and  the  plug  must  be  quickly  replaced  and 
covered  with  a  caoutchouc  cap.  If  collected  from  a  tap,  the  water 
should  first  be  allowed  to  run  for  a  few  minutes,  and  the  sample 
should,  be  received  into  the  flask  without  the  neck  coming  into  contact 
with  the  tap.  From  a  reservoir  or  stream,  the  flasks  may  be  filled 
by  employing  a  sterilised  pipette.  During  transport  contact  between 
the  water  and  cotton-wool  plug  must  be  avoided,  and  if  likely  to 
occur  the  sample  must  be  collected  and  forwarded  in  a  Sternberg's 
bulb. 

Examination    ~by   Plate-cultivation. — The    apparatus    for    plate- 
cultivation  should  be  arranged  as  already  described.     Crushed  ice 


FIG.  74. — APPARATUS  FOR  ESTIMATING  THE  NUMBER  OF  COLONIES  IN  A 
PLATE-CULTIVATION. 

may  be  added  to  the  water  in  the  glass  dish  to  expedite  the  setting 
of  the  gelatine,  so  that  the  plate  may  be  transferred  as  quickly 
as  possible  to  the  damp  chamber.  The  caoutchouc  cap  is  removed 
from  the  flask,  and  the  cotton- wool  plug  singed  in  the  flame  to 
prevent  contamination  from  adventitious  germs  on  the  outside  of 
the  plug.  The  flask  is  then  held  slantingly  in  .the  hand,  and  the 
plug  twisted  out  and  retained  between  the  fingers.  With  a 
graduated  pipette  a  measured  quantity  (y1^  or  -^  cc.)  of  the  sample  is 
transferred  to  a  tube  of  liquefied  nutrient  gelatine,  and  the  plugs  of 
the  flask  and  of  the  tube  quickly  replaced.  If  the  water  is  very 
impure,  it  may  be  necessary  to  first  dilute  the  sample  with  sterilised 
water.  The  inoculated  tube  must  be  gently  inclined  backwards  and 
forwards,  and  rolled  as  already  explained,  to  distribute  the  germs 
throughout  the  gelatine,  and  the  gelatine  finally  poured  on  a  plate. 
When  the  gelatine  has  set,  the  plate  is  transferred  to  a  damp 
chamber,  which  should  be  carefully  labelled  and  set  aside  in  a  place 


EXAMINATION   OF   AIR,    SOIL,    AND   WATER.  147 

of  moderate  temperature.  In  about  two  or  three  days  the  cultivation 
may  be  examined.  In  some  cases  the  colonies  may  be  counted  at 
once  ;  more  frequently  they  are  so  numerous  that  the  plate  must 
be  placed  on  a  dark  background,  and  a  special  process  resorted  to. 
A  glass  plate,  ruled  by  horizontal  and  vertical  lines  into  centimetre 
.squares,  some  of  which  are  again  subdivided  into  ninths,  is  so 
arranged  on  a  wooden  frame  that  it  can  cover  the  nutrient -gelatine 
plate  without  touching  it  (Fig.  74).  A  lens  is  used  to  assist  in  dis- 
covering minute  colonies.  If  then  the  colonies  are  very  numerous, 
the  number  in  some  small  division  is  counted,  if  less  in  some  large 
one,  and  an  average  is  obtained  from  which  the  number  of  colonies 
on  the  entire  surface  is  calculated.  A  separate  calculation  of  the 
liquefied  colonies  should  be  also  made,  and  their  number,  as  well 
as  the  total  number  of  colonies  present  in  1  cc.  of  the  sample, 
recorded.  Any  peculiar  macroscopical  appearances,  colours,  etc., 
should  be  noted,  and  then  the  microscopical  appearances  of  the 
colonies  studied.  Lastly,  examination  of  the  individual  organisms 
should  be  made  by  cover-glass  preparations,  and  by  inoculation  of 
nutrient  gelatine,  potatoes,  and  other  media. 

Instead  of  plates,  Petri's  dishes  may  be  used  both  for  gelatine 
.and  agar-agar  cultivations. 


FIG.  75. — ESM  ARCH'S  ROLL-CULTURE. 

.«,  India-rubber  caps;  b  b  b,  longitudinal  line  drawn  on  the  tube;  c,c,c,  transverse 
lines  for  counting  colonies  (FRANKLAND). 

Another  plan  is  to  take  a  measured  quantity  of  the  sample  of 
water  and  prepare  a  roll-culture,  using  a  large-sized  test-tube 
(Fig.  75).  The  colonies  can  be  counted  with  the  aid  of  a  lens  (Fig.  76). 
Microscopical  preparations  and  sub-cultures  can  be  made  from  the 
colonies,  and  the  anaerobic  bacteria  can  be  examined  by  Frankel's 
modification  of  this  method  (p.  131). 

A  drop  of  the  sample  of  water  may  also  be  added  to  liquefied 
nutrient  gelatine  in  a  test-tube,  the  organisms  distributed,  and  the 
gelatine  allowed  to  solidify  in  the  tube.  A  rough  comparison  of 
water  samples  may  be  made  in  this  way. 

Microscopic  Examination. — A  drop  of  the  water  may  be  mounted 
and  examined  without  staining ;  or  allowed  to  evaporate  on  a  cover- 


148  BACTERIOLOGY. 

glass,  which  is  then  passed  through  the  flame,  and  stained  in  the- 
usual  manner. 

Parietti's  Method. — As  typhoid  fever  bacilli  are  apt  to  be 
crowded  out  by  more  rapidly  growing  micro-organisms,  some  method 
had  to  be  devised  for  restraining  the  growth  of  the  latter,  and 
Chantemesse  and  Widal  suggested  the  use  of  carbolic  acid.  Parietti 
put  this  into  practice  by  the  method  he  introduced.  This  consists  in 
adding  to  tubes  of  broth  about  five  drops  of  a  mixture  composed  of 
sterilised  water  (100  parts),  hydrochloric  acid  (4  parts),  and  carbolic 
acid  (5  parts).  The  tubes  are  first  tested  by  incubation,  and  are 
then  ready  for  use.  A  few  drops  of  the  suspected  water  are  added 


FIG.  76.— APPARATUS  FOR  COUNTING  COLONIES  IN  A  ROLL  CULTURE. 

to  the  broth,  and  if  it  becomes  turbid  in  a  day  or  two  the  typhoid1 
fever  bacillus  is  present  in  the  form  of  a  pure- cult  tire. 

An  excess  of  bacteria  in  a  fresh  sample  indicates  an  excess  of 
organic  matter,  and  points  to  possible  contamination  with  sewage. 
Where  there  is  such  contamination  we  are  very  likely  to  find 
pathogenic  bacteria ;  and  moreover  impure  water  is  a  constant 
source  of  danger,  for  if  the  contagia  of  infectious  diseases  ar& 
introduced  they  will  retain  their  vitality  in  such  water  for  a  long 
period,  and  will  in  some  cases  even  multiply,  whereas  the  same 
organisms  introduced  into  pure  water  would  in  a  short  time  perish. 

The  actual  number  of  bacteria  in  water  is  not  of  very  great 
importance,  and  it  must  be  remembered  that  if  a  sample  is  set  aside 
for  a  few  days  there  will  be  an  enormous  increase  in  the  number 
of  bacteria  present ;  but  in  dealing  with  perfectly  fresh  samples  it 


EXAMINATION   OF   AIR,   SOIL,   AND   WATER.  149 

may  be  said  that  water  containing  less  than  100  bacteria  to  the 
cubic  centimetre  is  very  pure  water.  Water  containing  1,000  or 
more  should  be  filtered.  Water  containing  100,000  to  1,000,000 
is  contaminated  with  surface  water  or  sewage.  It  is  necessary  to 
bear  in  mind  that  in  typhoid  fever  and  Asiatic  cholera  the  excreta 
contain  the  bacteria  in  great  numbers,  and  wells  and  streams 
receiving  surface  water  may  be  contaminated  in  various  ways. 
The  cholera  bacillus  dies  as  a  rule  quickly  in  distilled  water,  while 
it  preserves  its  vitality  for  a  long  time  in  water  of  a  bad  quality. 

It  is  necessary  to  lay  stress  upon  the  fact  that  a  bacterio- 
logical analysis  may  show  the  presence  of  pathogenic  bacteria  when 
their  detection  is  not  possible  by  any  other  means.  They  may 
be  present  in  water  in  such  small  numbers  that  no  chemical 
analysis  would  detect  any  contamination,  but  as  they  are  living 
organisms  capable  of  increasing  in  a  suitable  environment,  they  can 
readily  be  discovered  by  bacteriological  methods. 

The  examination  of  rain  water,  drinking  water,  tap  water,  sea 
water,  various  liquids  and  infusions,  by  these  methods,  opens  up 
a  wide  field  for  research.  Pettenkofer  has  shown  that  impregna- 
tion of  water  containing  many  bacteria  with  carbonic  acid  diminishes 
the  number  of  the  latter.  The  examination  of  waters  before  and 
after  filtration,  or  after  addition  of  chemical  substances,  are  matters 
which  require  further  investigation,  though  a  great  deal  of  work  has 
already  been  accomplished.  The  reader  will  find  in  Micro-organisms 
in  Water  by  P.  and  G.  Frankland,  a  very  complete  account  of  this 
subject  with  valuable  analytical  tables. 


CHAPTER   XII. 

PHOTOGRAPHY    OF   BACTERIA. 

THE  production  of  pictures  of  microscopic  objects  by  photographic 
means  was  attempted  at  an  early  date.  Some  authorities  regard 
the  very  earliest  recorded  experiments  as  being  the  first  experi- 
ments alike  in  photography  and  micro-photography.  The  experi- 
ments of  Wedgwood  and  Sir  Humphry  Davy  were  embodied  in 
a  paper  read  before  the  Royal  Institution  in  1802.  They  obtained 
with  the  solar  microscope  impressions  upon  paper,  and  with  greater 
success  upon  white  leather,  though  the  results  were  transitory  when 
exposed  to  daylight. 

In  1816  Mcephore  Niepce  described  his  experiments  in  con- 
nection with  fixing  the  image  obtained  by  the  camera.  He  was 
at  first  only  able  to  obtain  negatives,  and  these  were  transitory. 
But,  after  joining  with  Daguerre,  who  had  been  experimenting 
in  the  same  direction,  a  process  was  invented  which  was  published 
in  1839  under  the  name  of  daguerreotype. 

This  invention,  and  the  rapid  improvements  which  followed, 
were  taken  advantage  of  by  Reade,  Donne,  Hodgson,  Kingsley,  and 
Talbot,  who  were  early  workers  in  the  field  of  micro-photography. 

So  early  as  1845  it  is  stated  that  Donne  produced  a  work 
illustrated  with  engravings  copied  from  daguerreotypes. 

Subsequently  this  interesting  branch  of  photography  was  taken 
up  by  many  in  France  and  Germany,  in  America,  and  in  England. 
Of  those  to  whom  we  are  indebted  for  the  literature  of  the  subject, 
and  for  many  improvements,  the  names  of  Wenham,  Dancer, 
Draper,  Maddox,  Shadbolt,  Redmayne,  Woodward,  Highley,  Deecke, 
Moitessier,  Gerlach,  Koch,  Sternberg,  Frankel,  Pfeiffer,  and  Pringle 
may  especially  be  mentioned. 

Of  these  workers  the  name  of  Woodward  stands  pre-eminently 
foremost.  His  skill  in  microscopical  manipulations,  combined 
with  access  to-  the  very  best  apparatus  and  objectives,  placed  at 

150 


PHOTOGRAPHY    OF    BACTERIA.  151 

his  disposal  in  the  Museum  at  Washington,  enabled  him  to  obtain 
photographs  of  diatoms  which  probably  have  never  been  surpassed. 

To  Koch  belongs  the  credit  of  being  the  first  to  extend  the 
application  of  micro-photography  to  the  delineation  of  bacteria. 
A  series  of  instructive  photographs  was  first  published  by  him 
in  1877.  These  were  photographs  of  cover-glass  preparations, 
and  all  admirably  illustrated  the  subjects  from  which  they  were 
taken ;  while  two,  showing  the  flagella  of  bacilli  and  spirilla, 
were  triumphs  in  this  new  departure. 

Lewis,  in  India,  was  one  of  the  first  to  illustrate  his  writings  on 
the  subject  of  micro-organisms  by  means  of  photographs. 

About  the  same  time  Sternberg,  in  America,  took  some  excellent 
photographs  of  bacteria.  Heliotypa  reproductions  of  these  were 
published  in  1884. 

Hauser  and  Van  Ermengem  and  many  other  bacteriologists 
successfully  resorted  to  photography  for  illustrating  their  researches, 
and  Frankel  and  Pfeiffer's,  and  Itzerott  and  Niemann's  atlases  of 
photographs  of  bacteria,  in  microscopical  specimens  and  cultivations, 
are  especially  worthy  of  mention. 

Opinions  have  differed  widely  as  to  the  merits  of  photographic 
illustrations.  Many,  taking  the  standpoint  solely  of  a  comparison 
with  drawings,  have  decried  their  use.  By  judging  from  such  a 
comparison  alone  the  real  value  of  photographs  may  be  lost  sight 
of.  On  the  other  hand,  many  who  have  looked  at  the  question 
from  all  sides,  have  been  led  to  value  even  a  defective  photograph 
more  than  an  ordinary  drawing. 

In  his  first  publication  on  this  subject,  Koch  strongly  advocated 
photography  on  the  ground  that  illustrations  would  then  be  as  true 
to  nature  as  possible.  The  photographs  which  accompanied  his 
paper  were  all  taken  from  preparations  of  bacteria  which  had 
been  made  from  blood,  cultivations,  or  infusions,  by  drying  a 
thin  layer  on  a  cover-glass  and  staining,  or  from  specimens  prepared 
in  the  same  way  but  left  unstained.  But  when,  having  committed 
himself  to  this  opinion,  Koch  attempted,  later,  to  photograph  the 
bacteria  in  animal  tissues,  he  was  led  to  modify  his  previous 
conclusion.  For  though  no  trouble  was  spared,  yet  disappointing 
results  were  met  with.  This  was  owing,  he  explains,  to  the  fact 
that  the  smallest  and  most  interesting  bacteria  can  only  be  made 
visible  in  animal  tissues  by  staining  them,  and  thus  obtaining  the 
advantage  of  colour. 

This  introduced  the  same  difficulties  which  are  met  with  in 
photographing  coloured  objects,  such  as  tapestry  and  oil  paintings. 


1 52  BACTERIOLOGY. 

As  these  difficulties  had  been  to  a  certain  extent  obviated  by  the 
use  of  eosin-collodion,  Koch  adopted  the  same  method  for  photo- 
graphing stained  bacteria.  By  the  use  of  eosin-collodion,  and  by 
shutting  off  portions  of  the  spectrum  by  coloured  glasses,  he 
succeeded  in  obtaining  photographs  of  bacteria  which  had  been 
stained  with  blue  and  red  aniline  dyes.  But,  owing  to  the  long 
exposure  which  was  necessary,  and  the  unavoidable  vibrations  of 
the  apparatus,  the  results  were  so  wanting  in  definition  that  they 
not  only  proved  unsatisfactory  as  substitutes  for  drawings,  but  did 
not  in  some  cases  give  any  evidence  of  what  was  to  be  seen  in  the 
preparations. 

Koch,  in  consequence,  stated  that  he  would  abstain  from 
publishing  photographic  illustrations  until  he  had  the  advantage 
of  improved  methods. 

We  find,  however,  in  spite  of  this,  that  in  1881  Koch  published 
a  series  of  reproductions  from  his  negatives  in  illustration  of  what 
could  be  accomplished  by  photography. 

Here  again  we  find  that  many  of  the  photographs  of  cover- 
glass  preparations  were  admirable,  but  those  of  tissue-sections  gave 
evidence  of  the  difficulties  Koch  encountered,  and  were  undoubtedly 
unsatisfactory  from  the  want  of  flatness  of  field,  some  of  the 
illustrations  recalling  rather  a  map  of  a  mountainous  country  than 
a  microscopical  preparation. 

In  consequence  of  the  difficulties  met  with  in  attempting  to 
photograph  bacteria  stained  with  the  aniline  dyes  most  commonly 
used,  Koch  recommended  that  the  preparations  should  be  stained 
brown,  pointing  out  as  his  reason  that,  though  the  bright  and 
concentrated  colour  of  the  red  and  blue  aniline  dyes  catches  the 
eye  far  more  readily  than  the  somewhat  sombre  brown  colours, 
yet  no  one  up  to  the  time  of  his  publication  had  succeeded  in 
obtaining  good  photographs  of  bacteria  which  had  been  stained 
either  blue  or  red,  and  mounted  in  Canada  balsam,  while  there  was 
no  difficulty  in  obtaining  photographic  representations  of  prepara- 
tions stained  yellow  or  brown. 

Though  this  stain  could  be  easily  employed  in  most  cover-glass 
preparations,  it  was  by  no  means  easy  to  obtain  a  good  differential 
stain  of  bacteria  in  the  tissues  by  employing  Bismarck  brown. 
An  attempt  was,  therefore,  made  to  photograph  preparations 
stained  blue  and  red  by  the  aid  of  the  dry-plate  process,  and  by 
interposing  glasses  of  suitable  tints.  After  many  fruitless  experi- 
ments this  method  had  to  be  abandoned,  arid  the  method  of  staining 
the  object  brown  was  adopted.  In  many  cases  this  gave  excellent 


PHOTOGRAPHY   OF   BACTERIA.  153 

results ;  in  others  again,  compared  with  the  results  of  staining 
with  blue  or  red  stains,  there  was  much  to  be  desired,  and  further 
improvement  was  needed. 

That  a  stain,  such  as  yellow  or  brown,  must  be  employed  which 
absorbs  the  blue  rays,  and  acts  on  the  sensitive  plate  like  black, 
which  absorbs  all  the  light,  constituted  the  first  condition  laid 
clown  by  Koch  as  an  essential  for  success.  It  was  further  pointed 
out  that  the  suitability  of  the  stain  could  be  ascertained  by  first 
passing  the  light,  to  illuminate  the  preparation,  through  a  solution 
of  ammonio- sulphate  of  copper,  under  which  condition  the  bacteria 
would  appear  black  on  a  blue  ground. 

The  second  condition  was,  that  sunlight  must  be  employed,  but 
that  direct  projection  upon  the  object  was  disadvantageous,  and  it 
must,  therefore,  be  diffused  by  the  interposition  of  one  or  more  plates 
of  ground  glass. 

Lastly,  an  illuminating  condenser  was  recommended,  of  such 
construction  that  the  diffused  sunlight  brightly  illuminates  the  object 
from  all  sides. 

Sternberg  encountered  the  same  difficulty  in  photographing  red, 
blue  or  violet  preparations,  while  he  produced  excellent  pictures  of 
preparations  stained  with  aniline  brown,  or  a  weak  solution  of  iodine 
(iodine  grs.  iij,  potassic  iodide  grs.  v,  distilled  water  grs.  200).  Thus 
the  results  of  a  large  number  of  attempts  to  photograph  the  tubercle 
bacillus  in  sputum,  only  ended  in  producing  such  extremely  faint 
impressions,  that  any  one  unacquainted  with  the  object  as  seen  under 
the  microscope  could  form  scarcely  any  idea  of  its  form  or  minute 
structure  with  even  an  accompanying  explanation  and  the  closest 
inspection  of  the  photograph. 

Dufrenne,  in  attempting  to  photograph  the  same  object  by  the 
ordinary  method,  found  the  plates  were  uniformly  acted  on,  or  the 
image  was  so  faint,  or  so  lacking  in  contrast,  that  they  were  useless 
for  obtaining  proofs  on  paper  or  glass.  By  interposing  green  glass 
between  the  objective  and  the  sensitive  plate,  so  that  the  red  rays 
were  absorbed,  while  the  green  rays  passed  through  and  acted  on 
the  plate,  he  states  that  better  results  were  obtained. 

The  work  of  Hauser  illustrated  the  great  value  of  photography 
in  the  production  of  pictures  of  impression-preparations  and  colonies 
in  nutrient  gelatine.  To  give  the  general  effect,  as  well  as  faithfully 
reproduce  the  minute  details  in  these  difficult  subjects  would  in  most 
cases  create  insurmountable  difficulties,  except  to  the  most  accom- 
plished draughtsman. 

Hauser   employed   Gerlach's   apparatus    and    Schleussner's   dry 


154  BACTERIOLOGY. 

plates,  and  obtained  the  illumination  by  means  of  a  small  incan- 
descent lamp,  which  gave  a  strong,  white  light,  with  three  or 
four  Bunsen  elements.  In  another  respect  Hauser's  results  were 
of  practical  value.  The  preparations  to  be  photographed  were 
stained  brown  as  recommended  by  Koch,  but  they  were  mounted  in 
the  ordinary  way  in  Canada  balsam.  The  objection  to  the  mounting 
medium  most  commonly  employed  was  thus  set  aside.  The  prevalent 
idea,  however,  that  the  preparations  must  be  stained  brown  was  still 
a  formidable  obstacle,  and  the  way  out  of  this  difficulty  was  clearly- 
shown  by  Yan  Ermengem's  photographs.  These  were  pictures  of 
comma-bacilli  which  had  been  stained  with  fuchsine  and  methyl 
violet.  These  photographs  afforded  the  first  practical  illustration 
of  the  value  of  isochromatic  plates  in  micro-photography  which  had 
been  previously  noted  by  Yan  Ermengem  in  1884,  and  their  intro- 
duction marks  a  distinct  era  in  the  progress  of  micro-photography. 

A  short  explanation  may  be  given  of  what  is  meant  by  isochro- 
matic, or  what  have  been  more  properly  termed  orthochromatic  dry 
plates.  The  difficulties  encountered  in  photographing  certain  stained 
preparations  have  been  mentioned.1  It  is  a  familiar  fact  that  in 
portraits,  blue  or  violet  comes  out  almost  or  quite  white,  while 
other  colours,  such  as  yellow,  are  represented  by  a  sombre  shade 
or  perhaps  black.  This  failure  in  correctly  translating  colours  is 
explained  by  the  want  of  equality  between  the  strength  of  the 
chemical  and  luminous  rays.  If  the  rays  of  the  spectrum  are  pro- 
jected upon  a  photographically  sensitive  surface,  the  greatest  effect 
is  found  to  take  place  at  the  violet  end.  In  other  words,  the  violet 
and  blue  rays  are  more  actinic  or  chemically  powerful,  while  the 
yellow  and  orange  have  scarcely  any  effect.  The  dyes  employed  in 
staining  give  corresponding  results  :  blue  and  violet  give  but  a  faint 
impression,  yellow  and  orange  a  black  picture.  These  results  are 
most  clearly  demonstrated  in  a  photograph  of  an  oil  painting  taken 
in  the  ordinary  way ;  and  they  led  to  experiments  being  made  which 
have  resulted  in  orthochroinatic  photography. 

The  effect  of  interposing  coloured  glasses  has  already  been 
referred  to.  It  was  found  later  that,  if  plates  were  coloured  yellow, 
e.g.,  with  turmeric,  the  blue  and  violet  rays  were  intercepted,  and 
their  actinism  reduced.  In  1881,  Tailfer  and  Clayton  produced  the 
so-called  isochromatic  plates.  The  emulsion  of  bromide  of  silver 
and  gelatine  was  stained  with  eosin,  and  it  was  claimed  that  colours 
would  be  represented  with  their  true  relative  intensity.  Chlorophyll 
and  other  stains  have  been  tried,  and  by  such  methods  the  ordinary 
gelatine  dry  plates  can  be  so  treated  that  they  will  reproduce 


PHOTOGRAPHY    OF   BACTERIA.  155 

various  colours,  according  to  their  relative  light  intensity,  and  thus 
be  rendered  iso-  or,  what  is  now  more  commonly  known  as,  ortho- 
chromatic. 

APPARATUS  AND  MATERIAL. 

Micro-photographic  Apparatus. — As  is  well  known,  various  forms 
of  apparatus  have  from  time  to  time  been  recommended  and  em- 
ployed by  different  workers. 

Many  use  the  microscope  in  a  vertical  position,  with  the  camera 
superposed  or  fitted  to  the  eye -piece  end  of  the  microscope  tube ; 
or  the  microscope  tube  may  be  screwed  off  from  the  body  of  the 
microscope,  and  a  pyramidal  camera  adjusted  in  its  place,  the  base  of 
the  pyramid  being  represented  by  the  ground  glass  screen. 

Others  again  prefer  that  the  microscope  and  camera  should  be 
arranged  horizontally. 

In  another  form  the  ordinary  microscope  is  dispensed  with,  the 
objective,  stage,  and  mirror  are  adapted  to  the  front  of  the  camera, 
and  provided  with  suitable  arrangements  for  holding  the  object, 
supporting  the  mirror,  and  adjusting  the  different  parts. 

Lastly,  the  camera  may  be  dispensed  with,  the  operating-room, 
which  must  be  rendered  impervious  to  light,  taking  its  place,  while 
the  image  is  projected  and  focussed  upon  a  ground  glass  screen, 
which  has  a  separate  support.* 

The  horizontal  position  affords  greater  stability  than  the  vertical, 
so  that  the  former  is  to  be  preferred.  The  vertical  model  with  the 
camera  fixed  to  the  microscope  is  particularly  to  be  avoided,  as  the 
weight  of  the  camera  bears  directly  upon  the  microscope,  and  must 
affect  the  fine  adjustment,  and  any  vibration  in  one  part  of  the 
apparatus  is  communicated  throughout. 

The  simplest  apparatus  consists  of  a  camera  fixed  upon  a  base- 
board four  or  five  feet  in  length,  upon  which  the  microscope  can  be 
clamped,  and  which  carries  also  a  lamp  and  a  bull's-eye  condenser 
(Fig.  77). 

Simplicity  and  economy  must  always  be  borne  in  mind  in 
recommending  any  apparatus  of  this  kind,  for  to  insist  upon  the 
necessity  of  a  very  elaborate  apparatus,  or  a  specially  fitted- up  room, 
or  that  a  special  room  should  be  built  with  windows  facing  in  a 
definite  direction,  will  in  most  cases  at  once  place  photography  beyond 
the  reach  of  those  who  might  otherwise  employ  it.  Yet  to  fulfil 

*  For  an  excellent  account  of  the  forms  of  apparatus  which  have  been 
employed  by  different  workers  the  reader  is  referred  to  the  section  on  Micro- 
photography  in  Beale's  How  to  work  with  the  Microscope. 


156 


BACTERIOLOGY. 


all  the  purposes  for  which  the  apparatus  may  be  required,  including 
the  employment  of  the  highest  powers,  and  also  that  one  may  be 
enabled  to  work  for  long  intervals  of  time  with  due  comfort,  an 
accurate  and  complete  apparatus  will  be  found  to  be  most  desirable. 
Though  most  preparations  will  admit  of  being  photographed 
when  the  stage  of  the  microscope  is  vertical,  yet  if  we  require  to 
photograph  micro-organisms  in  liquids,  or  colonies  upon  partially 
liquefied  gelatine,  the  apparatus  must  admit  of  being  placed  so  that 
the  stage  of  the  microscope  becomes  horizontal.  In  addition,  the 
apparatus  is  rendered  somewhat  complex  if  we  employ  powerful 


FIG.  77. — HORIZONTAL  MICRO-PHOTOGRAPHIC  APPARATUS. 


artificial  light.  Sunlight,  no  doubt,  is  the  best  and  cheapest,  but 
it  is  not  always  available,  especially  in  a  city  like  London;  and, 
moreover,  evenings  and  dull  days  will  probably  be  the  very  time 
which  can  be  best  spared  for  this  work.  We  must,  therefore,  fall 
back  upon  the  paraffine  lamp,  or  the  magnesium,  oxyhydrogen,  or 
electric  light. 

To  fulfil  all  these  conditions  Swift  has  constructed  an  apparatus 
under  the  author's  directions  (Fig.  78).  It  is  merely  a  modification 
of  the  ordinary  horizontal  model,  which  admits  of  being  readily  placed 


PHOTOGRAPHY    OF   BACTERIA. 


157 


in  the  vertical  position,  while  the  illumination  is  supplied  from  an 
oxyhydrogen  lantern. 

To  place  the  apparatus  in  the  vertical  position  two  small  hinged 


brackets,  at  the  end  distant  from  the  camera,  are  forced  up  with 
a  smart  blow  of  the  hand.  The  corresponding  ends  of  the  stretcher 
bars  are  dislodged  from  their  fittings,  and  allowed  to  descend  ;  when 


158  BACTERIOLOGY. 

horizontal,  the  opposite  extremities  of  the  bars  are  easily  released 
from  their  sockets.     The  leg  or  support  at  this  end  can  then   be 


FIG.  79. — REVERSIBLE  MICRO-PHOTOGRAPHIC  APPARATUS  ARRANGED  IN  THE 
VERTICAL  POSITION. 

turned  up  and  fixed  underneath  the  apparatus  by  a  button,  and 
the    end   of   the    apparatus   itself   gently  lowered   to   the   ground. 


PHOTOGRAPHY   OF    BACTERIA.  159 

A  hinged  end-piece  is  also  to  be  turned  out  to  increase  the  base  upon 
which  the  whole  apparatus  will  stand  when  raised  to  the  vertical. 
The  two-legged  support  at  the  opposite  end  of  the  apparatus  is 
next  worked  down  by  a  quick  thread  screw,  and  on  raising  the 
apparatus  to  the  vertical,  the  two-legged  support  drops  to  the 
ground,  and  assists  in  maintaining  the  stability  of  the  whole.  If 
it  is  thought  necessary,  a  simple  means  can  be  readily  devised  for 
clamping  the  apparatus,  in  either  position,  to  the  wall  of  the  room, 
so  as  to  eliminate  as  much  as  possible  all  chances  of  vibration.  A 
second  quick  thread  screw  moves  the  base-board  upon  which  the 
camera  and  central  sliding-board  are  mounted,  so  that  the  camera, 
microscope  and  lantern  can  be  raised  to  a  convenient  height  from 
the  ground. 

The  various  parts  of  this  apparatus  may  be  described  in 
detail. 

The  Microscope  and  its  Attachments. — It  is  most  essential  that 
the  microscope  should  be  perfectly  steady.  The  microscope  was  made 
by  Zeiss,  and  to  ensure  steadiness,  the  horse- shoe  foot  piece  tits  under 
a  projecting  ledge,  and  is  then  clamped  by  a  cross-piece,  so  that 
it  is  firmly  fixed. 

The  microscope  with  the  means  for  clamping  it  and  the  oxy- 
hydrogen  lantern  are  carried  upon  an  independent  sliding-board, 
which  admits  of  movement  to  or  from  the  camera.  The  sliding- board 
also  moves  upon  a  centre,  which  enables  the  microscope  to  be  turned 
out  from  the  median  line ;  in  fact,  to  be  turned  at  a  right  angle  to  the 
position  it  occupies  when  ready  for  the  exposure.  The  object  of  this 
contrivance  is  to  enable  the  operator  to  sit  down  by  the  side  of  the 
apparatus,  and  with  comfort  to  arrange  the  object  in  the  field  of  the 
microscope.  On  turning  the  microscope  back  into  the  median  line,  it 
is  fixed  in  the  optical  axis  of  the  apparatus  by  means  of  a  suitable 
stop.  The  sliding-board  is  provided  with  a  small  grooved  wheel 
receiving  an  endless  cord,  made  of  silk  or  fishing-line,  which  passes 
round  the  grooved,  milled  head  of  the  fine  adjustment  of  the 
microscope.  When  the  sliding-board  is  returned  to  the  median 
line  of  the  apparatus,  the  milled  wheel  connected  with  the  fine 
adjustment  impinges  upon  the  wheel  of  the  long  focussing  rod. 
The  latter  is  provided  with  an  india-rubber  tire,  which  grips  the 
teeth  of  the  milled  wheel,  and  thus  the  long  focussing  rod  is  placed 
in  connection  with  the  fine  adjustment  of  the  microscope. 

Illumination. — The  oxy-hydrogen  lamp  has  been  more  frequently 
employed  by  the  author  than  the  parafnne  lamp,  partly  on  account 
of  the  diminished  time  in  exposure,  especially  when  employing  very 


^-5w^ 

*?  KN. 

OF   TRR  X 

UNIVERSITY  ] 

°f  rTT^  ui 


160 


BACTERIOLOGY. 


PHOTOGRAPHY    OF   BACTERIA.  161 

high  powers ;  this  is  of  great  importance  where  there  is  likely  to  be 
vibration  from  passing  traffic.  With  rapid  plates  and  the  highest 
powers,  the  exposure  has  only  been  two  or  three  seconds,  whereas 
with  the  paraffine  lamp  it  may  vary  from  three  to  ten  minutes,  or 
even  longer. 

Walmsley    gives   the   following   table   for   exposures   with    the 
paraffine  lamp  : — 

1|  inch  objective        .          .          .          .          3  to  45  seconds. 

7        on 

3       »  ,,  .  .  .  .  l      ,,    v\J         ,, 

•=ir  i    . ,    3  minutes. 


l  9 

-§       »  »  .  .  .  .  Z 


i  4.          1 0 

10"  "  "  "  " 

The  illuminating  apparatus  represented  in  the  accompanying 
engraving  (Fig.  78)  consists  of  a  lantern  which  not  only  moves 
together  with  the  microscope  on  the  central  sliding-board,  but 
can  be  moved  independently  to  or  from  the  microscope,  and  be 
clamped  with  screws  at  the  requisite  distance  for  obtaining  the  best 
illumination.  It  is  provided  with  two  3-inch  condensing  lenses  of 
long  focus,  constructed  of  optical  glass,  which  is  much  whiter  than 
that  used  for  ordinary  lantern  condensers.  The  lime-cylinders  should 
be  of  the  hardest  and  best  quality,  as  they  give  a  more  actinic  light 
than  those  made  of  soft  lime.  The  "  Excelsior  "  lime-cylinders  are 
strongly  recommended.  They  are  supplied  in  hermetically  sealed 
tins  which  can  be  easily  opened  and  re-sealed,  so  that  a  cylinder  can 
be  taken  out  and  used,  and  the  rest  preserved  for  a  future  occasion. 
The  hydrogen  can  be  obtained  by  using  the  coal-gas  supplied  to  the 
house,  and  the  oxygen  should  be  supplied  preferably  in  a  compressed 
state  in  iron  bottles.  Not  only  are  the  bottles  much  less  cumbrous 
than  the  bags,  but  a  small  quantity  of  gas  can  be  used,  and  the 
residue  left  for  an  indefinite  time ;  moreover,  the  gas  is  always 
at  hand  to  be  turned  on  when  required.  On  the  other  hand,  the 
retention  of  unused  gas  in  bags  is  liable  to  cause  their  corrosion, 
owing,  it  is  believed,  to  impurities  which  are  carried  over  in  the 
manufacture  of  the  oxygen.  If  gas  is  not  laid  on  in  the  house,  then 
it  also  must  be  procured  in  a  compressed  state  in  bottles.  As  the 
blow-over  jet  is  recommended  on  account  of  its  safety,  the  bottles 
should  be  supplied  in  this  case  with  a  supplementary  valve.  It  is 
then  just  as  easy  and  free  from  danger  to  employ  the  compressed 
gas  as  it  is  to  make  use  of  the  house-supply. 

The  Camera. — A  long-focus,  half-plate  camera  is  mounted  upon 
a  sliding  platform.  This  admits  of  the  camera  being  pushed  up  to 

11 


162 


BACTERIOLOGY. 


the  microscope  when  it  is  in  the  long  axis  of  the  apparatus,  so  as  to 
make  a  light-tight  combination.  The  opening  which  is  filled  in  an 
ordinary  camera  by  the  lens  can  be  shut  off  by  means  of  an  internal 
shutter,  which  is  opened  and  closed  by  turning  a  screw  at  the  side 
of  the  camera.  The  dark-back  is  provided  with  plate-carriers,  so 
that  either  half,  quarter,  or  lantern-size  plates  can  be  employed.  It 
will  be  found  convenient  to  have  two  or  more  dark- backs,  so  that 
several  plates  may  be  exposed  without  rearranging  the  light  for 
each  exposure. 

Much  more  elaborate  and  expensive  micro- photographic  cameras 
have  been  constructed  by  Zeiss,  and  also  by  Swift.     The  latter  has 


FIG.  81.— PHOTOGRAPH  OF  AN  IMPRESSION  PREPARATION. 

carried  out  a  suggestion  made  by  Pringle  for  a  support  at  the 
ocular  end  (Fig.  80). 

The  Dark-room. — In  every  bacteriological  laboratory  there  should 
be  a  developing  room  provided  with  shelves,  gas,  water-tap,  and  sink, 
but  these  arrangements  are  not  absolutely  indispensable.  All  that 
is  essential  is  a  room  impervious  to  light ;  and  a  closet  or  cupboard, 
if  it  can  be  ventilated,  will  answer  perfectly  well,  with  a  jug  and 
basin  instead  of  the  tap  and  sink.  The  steam-steriliser  employed 
in  the  preparation  of  nutrient  media  for  cultivating  bacteria,  if  not 
required  at  the  time  for  such  purposes,  may  be  filled  to  the  brim 
with  water,  and  will  form  an  excellent  cistern  and  tap,  while  a  pail, 
or  small  sanitary  bin,  may  be  utilised  as  a  sink. 

Various  kinds  of  lamps  are  made  for  the  dark-room,  burning 


PHOTOGRAPHY   OF   BACTERIA.  163 

either  candles,  oil,  or  gas.  In  any  case,  the  light  must  pass  through 
two  thicknesses  of  ruby  glass. 

Dry  Plates. — A  small  supply  of  any  of  the  ordinary  plates  in 
the  market  may  be  procured  for  preliminary  trials  in  acquiring  a 
knowledge  of  the  processes  ;  but  to  overcome  the  difficulties  of  certain 
stained  preparations,  the  isochromatic  or  orthochromatic  plates  should 
be  used.  The  |  plate  will  be  found  to  be  the  most  suitable  size. 

There  are  numerous  formulae  for  the  requisite  solutions  for 
developing  and  fixing  the  negatives,  and  instructions  are  usually 
enclosed  in  the  boxes  of  dry  plates,  but  it  is  best  to  abstain  from 
trying  a  number  of  different  formulae,  as  it  leads  to  a  great  expendi- 
ture of  time.  There  is  a  temptation  to  do  this,  it  being  supposed 
that  there  is  probably  some  great  advantage  in  one  formula  over 
another.  It  is  much  better  to  get  accustomed  to  the  behaviour 
under  different  exposures  of  one,  or  perhaps  two  methods. 

In  France  the  iron  developer  is  much  in  vogue,  and  is  recom- 
mended by  Tailfer  and  Clayton  for  use  with  their  isochromatic 
plates.  It  has  the  advantage  of  great  simplicity  in  the  mode  of 
employment,  and,  therefore,  is  very  suitable  for  a  beginner.  In 
England,  on  the  other  hand,  the  alkaline  developer  is  very 
much  used,  as  it  gives  more  command  over  the  plate,  enabling  the 
photographer  more  fully  to  compensate  for  incorrect  exposure. 

It  is  very  desirable  before  attempting  to  take  photographs  with 
the  microscope  to  learn  how  to  take  photographs  with  an  ordinary 
landscape  camera,  and  to  get  thoroughly  accustomed  to  the  use  of 
some  good  developer,  so  that  mistakes  may  be  corrected  and  the 
clearest  and  sharpest  negatives  obtained. 

PRACTICAL  MANIPULATION. 

Arrangement  of  Apparatus. — For  working  with  the  paraffiiie 
lamp,  the  mode  of  procedure  is,  as  regards  the  illumination,  briefly 
as  follows.  The  sub-stage  condenser  is  dispensed  with  when  a 
low  power  is  employed,  as  well  as  the  mirror,  and  the  lamp  is 
so  placed  that  the  image  of  the  flat  of  the  flame  appears  accurately 
in  the  centre  of  the  field  of  the  microscope.  A  bull's-eye  condenser 
is  then  interposed,  so  that  the  image  of  the  flame  disappears,  and 
the  whole  field  is  equally  illuminated.  With  high  powers  the 
sub-stage  achromatic  condenser  is  necessary,  and  a  more  intense 
illumination  is  obtained  by  using  the  flame  edgewise.  In  using  a 
low  power  with  the  oxyhydrogen  light,  the  lantern  is  withdrawn 
some  little  distance  from  the  microscope,  and  the  top  combination 
of  the  achromatic  condenser  removed. 


164  BACTERIOLOGY. 

It  is  best  to  begin  with  the  use  of  a  low  power,  and  a  trial 
object,  such  as  the  blow-fly's  tongue,  spine  of  Echinus,  or  trachea 
of  silkworm. 

In  order  to  explain  the  management  of  the  apparatus  (as 
represented  in  Fig.  78)  the  steps  in  the  arrangement  of  the 
apparatus  and  exposure  of  the  plate  will  be  described  in  detail 
for  the  employment  of  a  high  power  and  the  oxyhydrogen  light. 
The  solutions  being  ready  for  use,  it  is  proposed  to  take  a  photograph 
of  tubercle  bacilli  in  sputum,  with  a  -f%  apochromatic  oil-immersion 
objective.  The  first  point  to  claim  attention  is  the  arrangement 
of  the  light.  Having  lighted  the  gas  at  the  hydrogen  jet,  the 
lime-cylinder  should  be  revolved  until  heated  equally  all  round. 
The  oxygen  is  then  carefully  turned  on  until  only  a  small  spot 
of  incandescence  is  produced.  The  central  sliding-board  is  turned 
out,  a  low  power  screwed  on  to  the  microscope,  and  the  image  of 
the  bright  spot  focussed  and  accurately  centered.  To  protect  the 
sight,  an  eye-pieca  provided  with  a  smoked  glass  shade  is  used. 
The  immersion  objective  is  then  substituted  for  the  low  power, 
and  the  oxygen  turned  on  until  the  right  admixture  of  gas  is 
obtained  to  produce  a  brilliant  illumination.  It  is  well  at  this 
stage  to  sit  down  to  focus  the  selected  object,  and  to  spend  some 
little  time  in  searching  for  the  most  characteristic  part  of  the 
specimen  to  be  photographed.  This  being  decided  upon,  the  eye- 
piece is  carefully  withdrawn,  and  the  central  sliding-board  rotated 
back  into  the  median  line.  To  make  a  light-tight  connection 
between  the  camera  and  the  microscope,  the  camera  is  pushed  up 
until  a  velvet-lined  tube,  which  occupies  the  position  of  the  lenses 
of  an  ordinary  camera,  is  enclosed  within  a  short  wide  tube  which 
is  adapted  to  the  eye-piec3  end  of  the  microscope. 

On  opening  the  camera-shutter  the  image  will  be  projected  upon 
the  ground  glass  screen  of  the  camera.  It  is  necessary,  however, 
to  obtain  the  exact  focus,  and  to  effect  this  the  ground-glass 
screen  is  turned  away,  and  the  dark-back  with  a  piece  of  plain 
glass  is  substituted.  Here  again  time  may  be  well  spent  in 
getting  the  sharpest  image,  with  the  aid  of  a  focussing  glass  of 
proper  focal  length. 

The  greatest  delicacy  in  manipulation  is  necessary,  as  in  working 
with  such  high  powers  a  turn  too  much  of  the  fine  adjustment  will 
cause  the  image  to  vanish.  Having  determined  the  best  visual 
focus,  which  will  be  found  with  the  high-power  objectives  of  most 
makers  to  correspond  with  the  chemical  focus,  the  dark-back  must 
be  cautiously  removed,  to  prevent  any  vibration,  and  the  plain 


PHOTOGRAPHY    OF   BACTERIA.  165 

glass  replaced  by  a  sensitive  plate.  To  effect  this  change,  the 
operator  retires  to  the  dark-room,  and  opens  a  box  of  plates  with 
as  little  exposure  to  the  red  light  as  possible.  Having  removed 
a  plate,  it  is  necessary  to  ascertain  which  is  the  sensitive  side* 
This  may  be  done  by  momentarily  exposing  it  to  the  red  light, 
and  seeing  which  is  the  sensitive  side  by  the  dull  appearance  of 
the  film.  A  less  satisfactory  way  is  to  moisten  the  tip  of  a 
finger,  and  press  it  at  one  corner  of  the  plate.  The  film  side  will 
be  recognised  by  imparting  a  sticky  sensation.  The  film  must  be 
dusted  with  a  camel's- hair  brush,  as  well  as  the  dark- back,  and 
the  plate  is  placed  film-side  downwards  in  the  dark- back,  which 
is  then  securely  closed. 

Care  should  be  taken  that  the  plates  then  remaining  in  the  box 
are  packed  away  before  light  is  admitted  to  the  dark-room. 

Exposure  of  the  Plate. — On  returning  to  the  apparatus,  the 
camera-shutter  is  closed.  Then  the  dark-back  is  gently  slid  into 
its  place,  and  its  slide  withdrawn.  A  few  moments  are  allowed 
to  elapse,  so  that  tne  least  possible  vibration,  which  might  be 
caused  by  inserting  the  dark- back,  has  had  time  to  cease,  and  all 
is  ready  for  the  exposure. 

In  the  case  of  the  object  we  have  selected,  three  seconds  will 
probably  be  the  exposure  required.  This  is  done  by  opening  and 
closing  the  earner  a -shutter  with  one  hand,  while  a  watch  can  be 
held  in  the  other.  The  slide  of  the  dark-back  is  then  carefully 
closed,  and  the  plate  is  ready  to  be  carried  off  to  the  developing 
room. 

As  the  light  will  not  be  again  required  until  the  next  exposure, 
the  oxygen  must  be  turned  off,  while  the  coal-gas  may  be  allowed 
to  play  over  the  lime. 

Development  and  Fixation  of  the  Image. — It  is  well  to  be 
systematic,  and  *  therefore,  before  the  plate  is  taken  out  of  the 
dark-back,  light  is  admitted  to  the  dark-room,  and  everything 
arranged  so  that  the  position  of  the  trays  and  bottles  may  be 
remembered  in  the  dark.  First,  let  the  ruby  lamp  be  lit,  place 
two  dishes  or  trays  close  by,  and  a  row  of  four  dishes  within  easy 
reach.  Pour  out  some  fixing  solution  in  the  first  porcelain  dish, 
alum  in  No.  2,  and  water  in  Nos.  3  and  4.  Put  the  necessary 
quantity  of  "  pyro "  solution  into  the  glass  measure,  and  place 
it  with  the  ammonia  drop-bottle  in  front  of  the  ruby  light. 
Then,  when  all  light  except  that  from  the  ruby  lantern  has  been 
excluded,  everything  is  ready  to  commence  the  development  of  the 
plate. 


166  BACTERIOLOGY. 

Opening  the  dark-back,  the  plate  may  be  turned  out  on  to  the 
palm  of  the  hand.  The  film  side  is  then  uppermost,  and  the  plate  is 
to  be  transferred  in  the  same  position  to  a  tray,  and  covered  with 
water.  This  is  to  soak  the  film  and  obtain  an  equal  action  of 
the  developer  ;  or  the  solution  of  fresh  pyro  may  be  poured  on  to 
the  plate  without  previous  soaking,  if  the  flow  is  uniform,  and  the 
formation  of  bubbles  avoided.  In  the  first  case  the  water  is  run  off 
and  the  pyro  allowed  to  flow  evenly  over  the  plate.  To  protect  the 
plate  from  prolonged  exposure  to  the  ruby  light,  a  second  tray  may 
be  inverted  over  it,  or  the  developing  tray  covered  with  a  piece 
of  card-board.  Gently  rock  the  tray  for  a  minute  or  so,  then 
to  a  few  drops  of  ammonia  in  a  measuring  glass  add  the  pyro 
from  the  developing  tray,  and  pour  the  mixture  back  again 
over  the  plate.  After  again  gently  rocking  the  tray  for  a  few 
minutes,  more  ammonia  is  added  by  drops  in  the  same  way. 
If  the  exposure  has  been  properly  timed, — and  the  time  necessary 
must  be  ascertained  by  trial  for  each  preparation, — the  image  will 
gradually  begin  to  appear,  and  the  action  must  be  allowed  to 
continue  until  sufficient  density  has  been  obtained.  To  determine 
this  requires  some  experience.  It  is  generally  recommended  to  take 
the  plate  out  of  the  tray  and  hold  it  for  a  moment,  film-side  towards 
the  operator,  in  front  of  the  ruby  light.  Though  the  plate  is  not 
nearly  so  sensitive  when  the  image  has  commenced  to  develop,  and 
there  is,  therefore,  not  the  same  danger  of  fogging,  a  safer  plan  is 
to  occasionally  turn  the  plate  film  downwards  in  the  tray,  and  when 
the  image  appears  on  the  back  the  development  will  be  found  to  be 
completed. 

With  such  a  preparation  as  tubercle  bacilli  in  sputum  it  is 
not  easy  to  trace  the  gradual  formation  of  the  image,  and  hence  the 
advantage  of  commencing  with  a  well-marked  object  such  as  the 
blow- fly's  tongue.  It  is  then  easy  to  watch  the  gradual  progress  of 
the  image.  The  bright  parts  or  high-lights  appear  first,  then 
gradually  the  half-tones,  or  less  brightly-lighted  parts,  and  lastly 
every  shade  except  the  deepest  shadows  is  represented.  When, 
however,  all  action  seems  to  have  ceased,  we  must  still  wait  until 
we  have  judged,  in  the  manner  already  described,  that  the  density 
is  sufficient.  This  being  determined,  we  pour  off  the  developing 
solution  and  thoroughly  wash  the  plate  with  water.  It  is  then 
ready  to  be  placed  in  dish  No.  1,  containing  "  hypo,"  and  here  it  must 
be  left  for  some  minutes  after  all  appearance  of  creaminess  has 
disappeared  from  the  back.  White  light  may  now  be  admitted,  the 
plate  removed  from  the  hypo  and  thoroughly  washed  under  the  tap, 


PHOTOGRAPHY    OF    BACTERIA.  167 

and  then  placed  in  dish  No.  2.  When  another  plate  is  ready  to 
take  its  place,  transfer  it  to  dish  No.  3,  and  then  to  No.  4,  and, 
after  a  good  final  washing  under  the  tap,  place  it  upon  a  rack 
to  dry.  If  there  is  any  tendency  for  the  film  to  detach  itself  from 
the  plate,  "  to  frill,"  the  alum  bath  must  be  used  before  fixing,  as 
well  as  after. 

Frilling  or  blistering  may  be  due  to  an  error  in  manufacture, 
and.  is  liable  to  occur  in  hot  weather,  or  when  using  a  developer  too 
strong  in  ammonia.  If  it  occurs  during  washing  or  fixing,  the  alum 
bath  must  be  employed  before  the  hypo.  Fogging,  or  the  appear- 
ance of  a  veil  over  the  plate,  may  arise  from  error  in  the  manu- 
facture, from  admission  of  extraneous  light,  from  over-exposure, 
or  from  prolonged  exposure  to  the  ruby  light  during  development. 
Care  must  be  taken  that  the  camera  and  dark-room  are  light-tight. 
Crystallisation,  or  powdery  deposit,  upon  the  negative  when  dry,  is 
due  to  insufficient  washing  out  of  the  hyposulphite  of  soda.  Thin- 
ness of  the  image,  or  want  of  density,  may  be  due  to  insufficient 
development,  too  weak  a  developer,  or  too  short  or  too  long  an 
exposure.  Too  great  density  results  from  too  long  immersion  in  the 
developer. 

Spots  may  sometimes  occur  upon  the  negatives.  They  may  be 
caused  by  dust  upon  the  plate  or  by  air  bubbles  in  the  developer. 

In  the  text-books  of  photography  full  accounts  of  failures  will  be 
found,  their  causes  and  prevention ;  but  it  will  be  advantageous 
when  these  difficulties  are  encountered  to  take  the  negatives  to  a 
skilled  photographer  and  get  advice  upon  them.  It  is  necessary 
to  persevere,  and  not  be  disheartened  if  several  negatives  have  to 
be  made  of  a  preparation  before  a  successful  result  is  obtained. 

It  may  here  be  remarked  that  the  beginner  will  far  more 
rapidly  learn  the  technique  if  he  avail  himself  of  a  practical  demon- 
stration from  a  photographer.  When  he  has  learnt  to  obtain  suc- 
cessful negatives,  if  he  prefer  silver  prints,  and  time  is  an  object,  it 
will  be  found  to  be  true  economy  to  get  the  printing  and  mounting 
done  by  a  professional  photographer.  The  credit  of  a  successful 
photograph  of  bacteria  is  due  to  the  bacteriologist  who  prepares 
the  microscopical  specimen  and  obtains  the  negative. 

Determination  of  the  Amplification. — The  amplification  varies 
not  only  with  the  objective  employed,  but  with  the  distance  of 
the  focussing  screen  from  the  object.  In  order  to  ascertain  the 
amplification  afforded  by  a  certain  objective  at  a  certain  distance,  a 
photograph  should  be  taken,  under  the  same  conditions,  of  the  lines  of 
a  micrometer  slide.  It  is  easy  then  to  calculate  the  amplification 


168 


BACTERIOLOGY. 


obtained  in  the  micro-photograph  ;  supposing,  for  example,  in 
the  micro- photograph  the  lines  which  are  i^Vrr  "1C^  apart  are 
delineated  1  inch  apart,  the  magnifying  power  must  be  1,000 
diameters.  Moreover,  having  thus  ascertained  the  amplification, 
we  can  accurately  compute  from  the  photograph  the  size  of  the 
objects  taken. 

Value  of  Photographs. — It  is  not  necessary  to  compare  the 
relative  merits  of  diagrams  and  photographs.  Diagrams  which  do 
not  purport  to  be  accurate  representations,  but  are 
intentionally  the  means  for  simplifying  instruction, 
will  always  be  valuable,  even  if  we  have  the 
original  preparations  under  the  microscope  before 
us.  We  must  consider  the  relative  merits  of 
photographs  and  of  drawings  which  purport  to  be 
exact  representations  of  what  is  seen  under  the 
microscope.  Thus  in  the  case  of  micro-organisms, 
when  their  biological  characters  are  studied  under 
low  powers  of  the  microscope,  photographs  are 
preferable  because  they  give  a  more  faithful  re- 
presentation. At  the  same  time,  apart  from  this 
comparative  value,  we  must  not  lose  sight  of  the 
actual  value  of  photography  in  placing  within  the 
reach  of  the  student  or  investigator,  who  may  not 
be  a  draughtsman,  a  most  valuable  means  for 
illustrating  all  kinds  of  preparations. 

For  double-stained  or  triple -stained  tissue  pre- 
parations an    accurately  coloured  drawing  leaves 
CULTIVATION     OF  little  to  be  desired  ;  but  if  we  reproduce  the  same 
BACILLUS        AN-   \yy  a  woocl  engraving,  and  so  lose  the  advantage 
of   the   coloured  picture,  which  is  instructive   in 
indicating  the  method  of  staining,  a  photograph  will,  in  many  cases, 
be  far  more  satisfactory. 

When  we  have  to  deal  with  the  growth  of  bacteria  en  masse, 
as  in  test-tube  and  plate- cultivations,  with  colonies  as  seen  under 
a  low  power  of  the  microscope,  and  with  impression-prepara- 
tions both  under  low  and  high  powers,  unless  the  bacteriologist 
is  a  most  accomplished  draughtsman  as  well  as  an  accurate  and 
reliable  observer,  photography  undoubtedly  affords  the  best  mode 
of  illustration.  The  apparatus  being  ready  and  at  hand,  a  negative 
can  be  produced  in  a  few  minutes  of  a  preparation  which,  from  the 
amount  of  detail  it  contains,  would  take  perhaps  several  hours  to 
draw  and  colour.  From  that  negative  any  number  of  facsimiles  can 


FIG.     82.  —  PHOTO- 
GRAPH       OF 


PHOTOGRAPHY    OF   BACTERIA.  169 

be  obtained,  whereas  an  original  drawing,  even  in  the  best  hands, 
if  cut  on  wood  or  lithographed,  is  almost  certain  to  fall  short  of 
being  an  exact  copy. 

With  regard  to  individual  bacteria,  the  result  is  more  satisfactory 
in  many  cases  than  a  drawing ;  for  there  is  the  advantage  of  being 
absolutely  certain  that  any  particular  structure,  form,  or  shape 
which  may  be  represented  is  actually  what  exists,  and  not  what 
may  have  been  evolved  by  unconscious  bias  in  the  mind  of  the 
observer.  Many  illustrations  might  be  given  of  this.  Thus  Lewis, 
who  was  a  most  conscientious  observer,  published  an  account  of 
organisms  in  the  blood  of  rats  in  India,  and  illustrated  it  with  a 
wood  engraving  and  with  micro-photographs.  The  identity  of  the 
organisms  which  were  found  in  the  common  brown  rat  of  this 
country  was  established  much  more  readily  from  •  these  photographs 
than  from  the  wood  engraving  or  the  description  in  the  letterpress. 

A  micro-organism,  even  under  the  highest  powers,  appears  as 
so  minute  an  object  that  to  represent  it  in  a  drawing  requires  a 
very  delicate  touch,  and  it  is  only  too  easy  to  make  a  picture  which 
gives  an  erroneous  impression  to  those  who  have  not  seen  the 
original.  If,  on  the  other  hand,  to  represent  the  object  more 
clearly  we  draw  an  enlarged  picture,  we  can  only  do  so  by  repre- 
senting what  we  think  the  object  would  be  like  if  it  could  be 
amplified  to  the  size  represented.  In  such  cases  a  photographic 
enlargement  is  certainly  more  valuable. 

Photography  enables  us  also  to  record  rapid  changes,  and  it 
is  possible  that  as  the  art  advances  we  may  find  that  the  film  is 
more  sensitive  than  the  human  retina,  and  brings  out  details  in 
bacteria  which  would  be  otherwise  unseen. 

Photographs  can  be  readily  transmitted  by  post,  and  when  we 
can  neither  make  a  great  number  of  preparations  to  illustrate 
!-orne  object,  nor  perhaps  be  able  to  go  to  the  expense  of  having  a 
drawing  reproduced,  this  method  will  be  of  value  in  enabling  others 
to  benefit  by  our  observations. 

The  author  is  convinced  that  if  the  employment  of  photography 
is  encouraged  in  bacteriological  and  other  research  laboratories  for 
depicting  microscopical  preparations  and  cultivations  of  bacteria,  the 
results  of  increasing  experience  and  practice  will  lead  to  its  being 
made  more  general  use  of  as  a  faithful  and  graphic  method,  valuable 
alike  for  class  demonstrations  and  for  illustrating  publications. 


PAET    II. 

ETIOLOGY  AND  PREVENTION  OF  INFECTIVE 
DISEASES. 


171 


CHAPTER   XIII. 

SUPPURATION,  PYAEMIA,  SEPTICAEMIA,  ERYSIPELAS. 

ABSCESS. 

WHEN  inflammation  is  followed  by  an  accumulation  of  leucocytes 
and  of  plasma  which  does  not  coagulate,  the  result  is  a  white  or 
creamy  liquid  called  pus,  and  when  the  surrounding  tissues  are 
involved  so  that  a  cavity  develops  containing  pus,  we  have  what 
is  termed  an  abscess.  The  almost  constant  association  of  bacteria 
with  the  production  of  pus  has  created  a  belief  that  they  are 
the  direct  cause  of  suppuration.  Ogston  found  micrococci  present 
in  all  acute  abscesses,  and  concluded  that'  acute  inflammation  was 
invariably  due  to  their  presence.  The  fact  that  inflammation 
occurs  more  frequently  in  the  external  tissues  of  the  body  is 
accordingly  explained  by  the  ready  entrance  of  bacteria  which 
are  in  the  air ;  and  suppuration  following  pericarditis,  pleurisy, 
and  other  conditions  in  which  air  is  excluded  is  attributed  to  the 
presence  of  pyogenic  cocci,  which  have  gained  access  by  the  blood 
stream.  There  is  no  pyogenic  organism  constantly  present,  but 
several  different  .species  of  bacteria  have  been  isolated  from  pus 
and  carefully  studied,  and  the  antiseptic  treatment  is  based  upon  the 
principle  of  excluding  bacteria  in  surgical  operations,  and  destroying 
any  which  may  have  previously  obtained  access  to  wounds  and 
broken  surfaces.  Inflamed  tissue  and  pus  form  a  most  suitable 
medium  for  the  growth  of  bacteria,  which  in  some  cases  are 
unquestionably  only  accidental  epiphytes. 

In  tuberculosis,  actinomycosis,  and  glanders,  pus  formation  may 
take  place  without  the  presence  of  pyogenic  cocci ;  and  it  is  generally 
believed  that  chemical  irritants,  such  as  croton  oil,  turpentine,  iodine, 
cadaverin,  and  tuberculin,  will  excite  the  formation  of  pus  in  the 
absence  of  bacteria,  although  Klemperer,  after  a  number  of  very 
careful  experiments,  maintains  that  no  genuine  pus  will  be  produced 
if  the  chemical  irritants  are  first  carefully  sterilised. 

173 


174 


INFECTIVE    DISEASES. 


The  bacteria  which  have  been  isolated  from  pus  include  :  — 
Staphylococcus  pyogenes  aureus,  albus,  and  citreus,  Staphylo- 
coccus  cereus  flavus  and  albus,  Streptococcus  pyogenes,  Micrococcus 
pyogenes  tenuis,  Micrococcus  pneumonia  crouposse,  Bacillus  pyo- 
cyaneus,  Bacillus  pyogenes  foetidus,  Micrococcus  tetragenus,  Bacillus 
intracellularis  meningitidis,  Gonococcus,  Bacillus  septicus  vesicse, 
Urobacillus  liquefaciens  septicus,  Bacillus  typhosus,  Bacillus  coli 
communis,  Bacillus  anthracis,  Bacillus  tuberculosis,  Bacillus  mallei, 
and  Actinomyces. 


FIG.  83. — SUPPURATION  OF  SUBCUTANEOUS  TISSUE. 

d,  Leucocyte  containing  micrococci ;  d\  leucocyte  with  pale  nucleus  showing 
necrosis ;  c,  fixed  connective  tissue  cells,  much  enlarged,  containing  several 
nuclei,  of  which  some  (n')  are  pale  and  necrotic ;  numerous  cocci,  diplococci, 
and  short  chains.  (CoRNiL  and  RANVIER.) 

Some  idea  of  the  distribution  of  the  bacteria  most  commonly 
occurring  in  pus  may  be  gathered  from  the  records  made  by  Passet 
and  by  Karlinski. 

Passet  examined  acute  abscesses,  and  found  Staphylococcus 
pyogenes  aureus  and  albus  in  11  cases,  Staphylococcus  pyogenes 
albus  alone  in  4,  Staphylococcus  pyogenes  albus  and  citreus  in  2, 
Streptococcus  pyogenes  alone  in  8,  Staphylococcus  pyogenes  albus 
and  Streptococcus  pyogenes  in  1,  and  Staphylococcus  pyogenes  albus 
and  citreus,  and  Streptococcus  pyogenes  in  1. 


SUPPURATION,    PY^MIA,    SEPTICAEMIA,    ERYSIPELAS.  175 

Karlinski  tabulated  his  cases  thus  : — 


. 

A 

§ 

.2 

1  § 

i  ^ 

0    * 

1  • 

"    CO 

3 

£'£• 

I 

t 

|| 

|| 

iq 

II 

5  3 

11 

jl 

| 

DISEASE. 

a 

"£>* 

^g 

'>>%> 

3  8> 

o  w1 

'§  8 

ll 

•<3 

|l 

A  fl 

ft 

CO 

II 

1! 

fl   CD 

| 

'§ 

PH 

"^ 

00  PK 

fr 

1 

Mastitis      

36 

22 

4 

4 

6 







— 

Subcutaneous  Abscess 

30 

10 

2 

8 

6 

2 

2 



— 

Phlegmon  ..... 

24 

_ 



24 



— 



— 

Furuncle    ..... 

20 

9 



10 



1 

— 



— 

Bubo  

17 

8 

1 

1 

7 







— 

Subperiosteal  Abscess 

16 

6 



10 









— 

Panaritium  Cutaneum 

16 

7 



9 









— 

Panaritium  Osseum   . 

10 

7 



3 





— 



— 

Dental  Abscess  .... 

10 

1 



4 

1 

3 

1 



— 

Horclcolviiii 

in 

i? 

4* 

Abscess  of  the  Middle  Ear 

1U 

4 

O 

2 

j 

_ 

2 

Carbuncle  ..... 

4 

2 



1 

1 



— 

4 

Osteomyelitis     .... 

3 

2 

— 

1 

— 

— 

— 

— 

— 

Total  

200 

82 

7 

55 

45 

6 

3 

2 

4 

PY^MIA  AND  SEPTICAEMIA. 

When  pyogenic  micrococci  get  access  to  the  blood  stream  they 
may  be  carried  into  distant  parts,  and  by  multiplying  produce  meta- 
static  abscesses  in  the  lymphatic  glands,  bones,  joints,  and  internal 
organs,  a  condition  which  is  recognised  as  pyaemia. 

If  there  is  a  general  invasion  of  the  blood  stream  by  micrococci, 
and  absorption  of  their  poisonous  products,  septicaemia  results,  and 
death  may  occur  before  the  development  of  any  secondary  lesions. 
When  septic  micro-organisms  multiply  locally,  and  their  chemical 
products  are  absorbed,  or  their  products  are  separated  from  putrid 
material  and  injected  into  the  circulation,  the  result  may  be  called 
sapraymia.  The  blood  in  septicaemia  contains  living  organisms,  and  is 
infective.  The  blood  in  sapraemia  contains  only  the  toxic  chemical 
products,  and  is  not  infective.  The  one  is  septic  infection  and  the 
other  septic  intoxication.  Pyaemia  may  follow  accidental  wounds, 
surgical  operations,  parturition,  acute  suppuration  of  bones,  scarlet 
fever,  typhoid  fever,  and  other  diseases. 

To  avoid  pyaemia  in  surgery  and  midwifery,  the  greatest  care 
must  be  taken  to  prevent  micro-organisms  from  being  conveyed  by 
instruments,  sponges,  bandages,  and  by  the  hands  of  the  surgeon 
or  the  obstetric  physician.  By  the  use  of  antiseptics  and  absolute 
cleanliness  the  chances  of  infection  are  reduced  to  a  minimum. 


176  INFECTIVE    DISEASES. 

Rosenbach  examined  six  cases  of  inetastatic  pyaemia :  Strepto- 
coccus pyogenes  was  found  five  times,  partly  in  the  blood  and  partly 
in  the  metastatic  deposits,  and  twice  in  company  with  Staphylococcus 
pyogenes  aureus. 

Baumgarten,  also,  found  Streptococcus  pyogenes  in  the  internal 
organs  in  pysemic  cases,  and  Eiselsberg  found  Streptococcus  pyogenes 
in  company  with  Staphylococcus  pyogenes  aureus  in  the  blood  of 
cases  of  septicaemia. 

Frankel  isolated  a  streptococcus  from  puerperal  fever,  which 
he  at  first  called  Streptococcus  puerperalis,  but  subsequently 
identified  with  Streptococcus  pyogenes.  These  researches  have 
been  confirmed  by  others.  Winkel  obtained  a  pure  cultivation  of 
a  streptococcus  from  the  blood  of  the  heart  in  a  case  of  puerperal 
peritonitis.  It  produced  erysipelatous  redness  when  inoculated 
in  the  rabbit's  ear,  and  in  form  arid  in  cultivation  was  similar  to 
the  streptococcus  in  erysipelas.  Gushing  also  found  Streptococcus 
pyogenes  associated  with  puerperal  infection.  The  cocci  were 
found  in  endometritis  diphtheritica  as  well  as  in  secondary  puerperal 
inflammation.  These  observations  were  still  further  confirmed  by 
Baumgarten,  and  Bumm  isolated  the  same  organism  in  puerperal 
mastitis. 

DESCRIPTION  OF  BACTERIA  IN  Pus. 

A  description  may  now  be  given  of  the  cocci  most  frequently 
found.  Staphylococcus  pyogenes  aureus  and  albus  and  Strepto- 
coccus pyogenes  and  Gonococcus  are  the  most  important  of  these. 
Staphylococcus  pyogenes  citreus,  cereus  albus  and  flavus,  are  pro- 
bably merely  epiphytic.  Micrococcus  tetragenus,  Micrococcus 
pyogenes  tenuis,  Bacillus  pyogenes  foetid  us,  Bacillus  pyocyarieus, 
Bacillus  coli  communis,  Bacillus  septicns  vesicse,  Urobacillus  lique- 
faciens  septicus,  arid  Bacillus  intracellularis  meningitidis  will  be 
described  fully  in  Part  III.  The  description  of  Actinomyces,  of 
Micrococcus  pneumonise  crouposse  and  of  the  bacilli  of  anthrax, 
tuberculosis,  glanders,  and  typhoid  fever,  will  be  found  in  other 
chapters  in  Part  II. 

Staphylococcus  pyogenes  aureus  (Rosenbach). — Yellow 
coccus  in  pus  (Ogston).  Cocci  singly,  in  pairs,  very  short  chains,  and 
irregular  masses.  Cultivated  on  nutrient  agar-agar,  an  orange - 
yellow  culture  develops,  looking  like  a  streak  made  with  oil  paint. 
One  variety  grows  on  nutrient  gelatine  without  liquefying  it ; 
another  produces  rapid  liquefaction,  and  the  growth  subsides  as 


SUPPURATION,    PYAEMIA,    SEFOCLEMIA,    ERYSIPELAS. 


177 


aii  orange -yellow  sediment.  On  potatoes  and  blood  serum  a  similar 
orange-yellow  culture  grows  luxuriantly.  They  may  also  form 
colourless  growths  in  sub-cultures,  and  are  then  indistinguishable 
from  Staphylococcus  pyogenes  albus.  The  cocci  do  not  cause  any 
septic  odour  in  pus,  nor  does  any  gas  develop.  Albumin  is  con- 
verted by  their  action  into  peptones. 
They  produce  rapid  ammoniacal  fer- 
mentation in  urine  (Shattock). 

The  micro-organisms  injected  into 
the  pleura  or  knee  of  a  rabbit  produce, 
as  a  rule,  a  fatal  result  on  the  following 
day  ;  but  if  it  survives  longer,  it  eventu- 
ally dies  of  severe  phlegmon.  If  injected 
into  the  knee  of  a  dog,  suppuration 
occurs,  followed  by  disintegration  of  the 
joint.  Injected  into  the  peritoneal 
cavity  of  animals,  they  set  up  perito- 
nitis, and  introduced  into  the  jugular 

vein  they  produce  septicaemia  and  death.  When  a  small  quantity  of 
a  cultivation  is  introduced  into  the  jugular  vein  after  previous  fracture 
or  contusion  of  the  bones  of  the  leg,  the  animal  dies  in  about  ten  days, 
and  abscesses  are  found  in  and  around  the  bones,  and  in  some  cases  in 

the  lungs  and  kidneys, 
and  the  cocci  are  found 
in  the  blood  and  pus. 
Gar  re  caused  sup- 


FIG.  84. — Pus  WITH  STAPHYLO- 
cocci,  x  800  (FLUGGE). 


wound  near  his  finger 
nail.  Bockhart  suffered 
from  several  pustules 
after  vaccinating  his 
arm  with  a  pure-culture 
suspended  in  salt  solu- 


himself  a  hypodermic 
FIG.  85.— SUBCUTANEOUS  TISSUE  OF  A  RABBIT  48  injectioii  of  a  pure- 

HOURS    AFTER  AN   INJECTION  OF  STAPHYLOCOCCI,   X  * 

«.»:>(>  (IiAt-.M«jAKTKN).  culture   and   produced 

an  abscess.  This  micro- 
organism is  practically  ubiquitous.  It  has  been  cultivated  from 
the  skin  and  mucous  membranes  and  secretions  of  healthy  persons, 
a  iul  it  occurs  in  the  air,  in  soil,  in  dust,  and  in  water,  and  in 

12 


178  INFECTIVE    DISEASES. 

association  with  suppuration,  pyaemia,  puerperal  fever,  and  acute 
osteomyelitis. 

Staphylococcus  pyogenes  albus  (Rosenbach).— Cocci  micro- 
scopically indistinguishable  from  the  above.  In  cultivations  also 
they  resemble  Staphylococcus  pyogenes  aureus,  but  the  growth 
consists  of  opaque  white  masses.  They,  as  a  rule,  liquefy  nutrient 
gelatine  very  rapidly,  and  subside  to  the  bottom  as  a  white  sediment ; 
more  rarely  they  liquefy  very  slowly ;  and  a  variety  has  also  been 
described  which  does  not  produce  any  liquefaction.  They  are  similar 
to  the  above-mentioned  in  their  pathogenic  action.  Pure-cultivations 
of  the  organism  were  obtained  from  a  case  of  acute  suppuration 
of  the  knee-joint. 

Staphylococcus  pyogenes  citreus  (Passet). — Cocci  singly,  in 
pairs,  very  short  chains,  and  irregular  masses.  If  cultivated  on 
nutrient  gelatine  or  nutrient  agar-agar,  a  sulphur  or  lemon-yellow 
growth  develops.  When  inoculated  under  the  skin  of  mice,  guinea- 
pigs,  or  rabbits,  an  abscess  forms  after  a  few  days,  from  which  a 
fresh  cultivation  of  the  micro-organism  can  be  obtained. 

Staphylococcus  cereus  albus  (Passet). — Cocci,  morphologic- 
ally similar  to  the  above,  but  distinguished  by  forming  on  nutrient 
gelatine  a  white,  slightly  shining  layer,  like  drops  of  stearine  or  wax, 
with  somewhat  thickened,  irregular  edges.  In  the  depth  of  gelatine 
they  form  a  greyish- white,  granular  thread.  In  plate-cultiva- 
tions, on  the  first  day,  white  points  are  observed,  which  spread 
themselves  out  on  the  surface  to  spots  of  1  to  2  mm.  When  culti- 
vated on  blood  serum  a  greyish-white,  slightly  shining  streak 
develops,  and  on  potatoes  the  cocci  form  a  layer  which  is  similarly 
coloured. 

Staphylococcus  cereus  flavus  (Passet). — Cocci  which  produce 
in  nutrient  jelly  a  growth  which,  at  first  white,  becomes  lemon- 
yellow,  somewhat  darker  in  colour  than  Staphylococcus  pyogenes 
citreus.  Microscopically  Staphylococcus  cereus  flavus  corresponds 
with  Staphylococcus  cereus  albus.  Inoculation  experiments  with 
both  kinds  give  negative  results. 

Streptococcus  pyogenes  (Rosenbach).~Cocci  occurring  singly, 
in  masses,  and  in  chains.  The  individual  cocci  are  small  spherical 
cells,  with  a  special  tendency  after  fission  for  the  resulting  elements 
to  remain  attached  to  each  other,  forming  chains  or  rosaries.  In 
cultures  on  solid  media  they  often  occur  in  the  form  of  staphylococci, 
but  in  liquid  cultures  there  may  be  a  few,  three  or  more  elements, 
linked  together ;  or  a  great  number,  forming  long  chains  which  may 
be  straight,  serpentine,  or  twisted. 


DESCRIPTION    OF    PLATE    IV. 
Streptococcus  Pyogenes. 

FIG.  1.— -From  a  cover-glass  preparation  of  pus  from  a  pyaemic  abscess. 
Stained  with  gentian-violet  by  the  method  of  Gram,  and  contrast-stained 
with  eosin.  x  1200.  Powell  and  Lealand's  apochromatic  -^  Horn.  imm. 
E.  P.  10. 

FIG.  2. — From  cover-glass  preparations  of  artificial  cultivations  of  the  strepto- 
coccus in  broth  and  in  milk  at  different  stages  of  growth,  x  1200.  Powell 
and  Lealand's  apocbromatic  ^  Horn.  imm.  E.  P.  10. 

Jn  these  preparations  there  is  a  great  diversity  in  size  and  form  of  the 
chains  and  their  component  elements.  In  the  drawing  examples  are 
figured  of  the  following: 

(a)  Branched  chains. 

(b)  Simple   chains    composed   of  elements  much   smaller  than  the 
average  size.  v 

(c)  Chains  with  spherical  and  spindle-shaped  elements  at  irregular 
intervals.     These  are  conspicuous  by  their  size,  and  are  sometimes 
terminal. 

(d  e)  Chains  in  which  the  elements  are  more  or  less  uniform  in  size. 
(/)  Complex  chains  with  elements  dividing  both  longitudinally  and 

transversely,  and  varying  considerably  in  size  in  different  lengths 

of  the  sam.e  chain. 


Plate  IV 


xrx 


.* 


Figl. 


../ 


Fig  2 


STREPTOCOCCUS  PYOGENES 


Vincent  Bnolta,  Day  *Son,,Iil)i. 


SUPPURATION,    PY^MIA,    SEPTICAEMIA,    ERYSIPELAS.  179 

The  individual  elements  composing  the  chains  will  be  found  to 
vary  considerably  in  size  :  here  and  there  in  a  preparation  will 
be  found  a  chain  composed  of  excessively  small  cocci,  in  another 
part  the  elements  will  be  all  on  a  larger  scale,  and  again  in 
another  part  they  will  be  peculiarly  conspicuous  on  account  of  their 
si/o.  So  great  is  the  diversity  in  the  size  of  the  cocci  in  some  of  the 
chains,  that  one  might  imagine  that  there  was  more  than  one  kind 
of  streptococcus  present  in  a  preparation,  until  on  examining  some 
of  the  longest  chains  it  is  observed  that  various  sizes  are  repre- 
sented in  different  lengths  of  the  same  chain.  Very  characteristic 
appearances  result  from  the  fact  that  the  cocci  enlarge  and  divide 
both  longitudinally  and  transversely  ;  and,  indeed,  the  largest,  for 
the  most  part,  clearly  show  a  division  in  two  directions,  resulting 
in  the  formation  of  tetrads.  In  addition  to  the  forms  resulting 
from  the  fission  of  the  cocci,  there  are  here  and  there  in  a  chain, 
and  sometimes  terminally,  larger  elements,  which  are  spherical, 
spindle-shaped,  or  in  the  form  of  a  lemon.  In  the  length  of  the 
chains,  as  in  the  size  of  the  individual  cocci,  there  is  usually  great 
diversity,  In  some  cases  they  are  composed  of  only  a  few,  three 
or  four  cocci;  in  others  eight,  ten,  or  twenty.  Here  and  there 
an  exquisite  rosary  will  extend  in  a  straight  line  across  the  field 
of  the  microscope,  or  be  twisted,  curved,  or  serpentine  ;  in  some 
preparations  twisted  or  entangled  strands  are  observed  which  are 
composed  of  several  hundred  elements.  Such  chains  will  be  found 
to  be  much  thicker  in  one  part  than  another.  Another  char- 
acteristic appearance  is  produced  by  separation  of  the  elements 
resulting  from  fission  in  the  long  direction  of  the  chain,  by  which 
lateral  twigs  or  branches  are  formed.  Another  character,  which 
is  very  striking,  may  be  seen  when  the  individuals  in  a  chain 
have  become  separated ;  an  unstained  or  faintly  stained  membrane 
may  be  found  bridging  across  the  interval.  This  will  become  still 
more  visible  in  preparations  contrast-stained  with  eosiu. 

In  plate-cultivations  the  appearances  of  the  colonies  are  not  very 
striking.  They  appear  to  the  naked  eye  after  three  or  four  days 
as  extremely  minute,  greyish-white,  translucent  dots,  which  under 
the  microscope  have  a  slightly  yellowish -brown  colour.  They  are 
finely  granular  and  well  defined.  They  do  not  liquefy  the  gelatine, 
and  after  several  weeks  may  not  exceed  the  size  of  a  pin's  head. 

If  the  surface  of  nutrient  gelatine  solidified  obliquely  be  traced 
over  once  or  twice  with  a  platinum  needle  bent  at  the  extremity 
into  a  little  hook  charged  with  the  cocci,  a  ribbon-shaped  film 
develops  in  two  or  three  days.  This  film  is  composed  of  minute, 


180  INFECTIVE    DISEASES. 

greyish-white,  translucent  dots  or  droplets,  which  can  be  more  easily 
recognised  with  the  aid  of  a  pocket  lens  (Fig.  87).  According  to 
the  number  of  organisms  sown  on  the  jelly,  the  dots  or  colonies 
may  be  completely  isolated,  or  form  a  more  or  less  continuous 
film.  The  film  by  reflected  light  has  an  iridescent  appearance  like 
mother-of-pearl,  but  has  a  bluish  or  bluish-grey  tint  by  transmitted 
light,  and  with  a  pocket  lens  appears  distinctly  brownish.  The 
gelatine  is  not  liquefied,  and  even  after  several  weeks  the  cultivation 
is  limited  to  the  inoculated  area,  and  the  individual  colonies  are,  as 
a  rule,  not  larger  than  pins'  heads.  In  gelatine-cultivations  of  the 
same  age,  but  kept  in  the  incubator  at  18°  C.,  the  colonies  get 
irregular  in  form,  especially  at  the  margin  of  the  film,  and  give  the 
growth  an  arborescent,  fringed,  or  serrated  appearance.  Cultivated 
on  the  oblique  surface  of  nutrient  agar-agar  at  37°  C.  the  growth  is 
very  similar,  forming  a  film  composed  of  minute  white  colonies 
like  grains  of  sand ;  but  the  film  appears  less  transparent,  is 
whiter,  and  the  colonies  have  a  greater  tendency  to  get  irregular 
in  form.  If  inoculated  with  one  tracing  of  the  needle  the  growth 
is  scanty,  but  tends  to  get  thicker  in  the  centre  than  towards  the 
margins,  which  may  have  a  terraced  appearance.  Inoculated  in 
the  depth  of  gelatine,  there  appears  after  a  day  or  two  at  18°  C.  a 
thread-like  growth  along  the  track  of  the  inoculating  needle.  This 
delicate  growth  is  found  on  examination  with  a  pocket  lens  to  consist 
of  a  linear  series  of  extremely  minute  granules.  In  a  few  days 
more,  the  beads  or  granules  become  more  marked,  but  even  after 
weeks,  the  cultivation  only  appears  like  a  string  of  minute,  white, 
compact,  globular  masses  or  grains.  In  broth  at  37°  C.  the  cocci  in 
twenty-four  hours  create  a  turbidity,  and  gradually  develop  beauti- 
ful chains  varying  in  length  according  to  the  age  of  the  cultivations. 
Even  in  forty-eight  hours  there  may  be  chains  of  eight,  ten,  twenty, 
or  a  hundred  elements.  After  a  few  days  the  growth  settles  down 
at  the  bottom  of  the  tube  in  the  form  of  a  white  deposit,  while  the 
supernatant  liquid  becomes  again  clear. 

Inoculated  subcutaneously  in  the  ear  of  rabbits,  they  produce  in. 
two  days  an  inflammatory  thickening  with  erysipelatous  redness, 
or  sometimes  suppuration. 

They  may  occur  in  vaccine  lymph,  as  the  result  of  con- 
tamination, and  Pfeiffer  suggested  that  before  calf  lymph  is 
employed  for  vaccination  it  should  be  tested  on  a  rabbit's  ear. 
If  in  two  days  no  rash  has  been  produced,  the  possibility  of  the 
presence  in  the  lymph  of  Streptococcus  pyogenes  or  erysipelatis 
is  excluded. 


SUPPURATION,   PY^MIA,   SEPTICAEMIA,    ERYSIPELAS.  181 

According  to  Fliigge  and  others,  after  subcutaneous  inoculation 
of  mice  with  a  small  quantity  of  a  cultivation,  there  is  no  result  in 
80  per  cent,  of  the  animals  experimented  upon.  Sometimes  there 
is  limited  pus  formation  at  the  seat  of  inoculation,  sometimes  the 
animals  die  without  any  very  striking  pathological  appearances. 

They  occur  in  abscesses,  pyaemia,  and  septicaemia,  and  are  often 
found  in  diseases  such  as  scarlet  fever  and  typhoid,  associated  with 
septic  complications.  They  have  been  isolated  from  air,  soil,  and 
water. 

The  streptococcus  found  in  erysipelas  agrees  in  description,  and 
is  merely  a  variety  of  Streptococcus  pyogenes.  It  has  been  definitely 
established  by  the  researches  of  Frankel  and  Freudenberg,  and  later 
by  those  of  the  author,  Raskin,  Prudden,  and  Bayard  Holmes, 
that  Streptococcus  pyogenes  is  frequently  found  in  scarlet  fever  and 
diphtheria,  and  in  other  diseases  associated  with  septic  complica- 
tions. The  author  has  isolated  Streptococcus  pyogenes  from  acute 
abscesses,  from  suppuration  after  surgical  operations,  from  pyaemia, 
from  pyaemia  after  scarlet  fever,  and  from  purulent  peritonitis. 
Some  of  these  cultures  have  been  kept  up  for  very  long  periods, 
extending  over  some  years,  so  that  opportunities  occurred  for  a 
complete  investigation  into  the  life  history  of  this  micro-organism. 
Variations  in  the  appearances  of  cultures  have  been  observed  when 
obtained  from  the  same  source.  A  number  of  cultures  from  pus 
were  prepared  011  gelatine  and  agar,  made  according  to  the  usual 
formula,  but  at  different  dates,  and,  therefore,  varying  slightly  in 
composition  and  quality.  Sub-cultures  were  also  started  in  nutrient 
gelatine  of  precisely  the  same  composition,  but  from  primary  cul- 
tures of  the  same  micro-organism  in  different  media — agar-agar, 
milk,  and  broth.  The  descriptions  of  the  streptococcus  hitherto 
published  were  then  found  to  be  inadequate.  The  different  cultures 
and  sub-cultures  presented  striking  variations  in  the  microscopical  and 
macroscopical  appearances.  Some  sub-cultures  on  gelatine,  for  exam- 
ple, exhibited  a  finely  dotted  appearance,  others  showed  every  variety 
in  the  size,  and  degree  of  opacity  of  the  colonies  (Fig.  89).  Cultures 
in  broth  also,  varied  in  appearance,  owing  to  slight  variation  in  the 
composition  of  the  medium,  to  slight  differences  of  temperature,  an«l 
other  conditions  difficult  to  determine.  The  addition  of  glycerine  to 
broth  materially  alters  the  appearance  of  the  culture.  It  was  con- 
clusively proved  that  minute  differences  arise  from  different  conditions 
of  the  cultivating  media.  The  author  was  led  to  study  exhaustively 
the  streptococcus  of  acute  suppuration  in  bo  vines.  Primary  cultures 
of  Streptococcus  pyogenes  from  man,  and  primary  cultures  from 


182  INFECTIVE    DISEASES. 

a  case  of  purulent  peritonitis  in  a  cow,  were  carried  through 
sub-cultures  under  exactly  similar  conditions.  -Cultivations  of  the 
Streptococcus  pyogenes  bovis  exhibited  variations  in  microscopical 
and  cultural  characters  which  were  even  more  marker!  than  in  the 
case  of  the  Streptococcus  pyogenes  hominis.  By  selecting  certain 
cultures  from  both  sources  there  was  a  striking  similarity  if  not 
identity  between  them,  but,  when  compared  under  exactly  identical 
conditions,  there  was  more  difference  in  cultural  characters  between 
the  Streptococcus  pyogenes  bovis  and  the  Streptococcus  pyogenes 
hominis  than  between  the  Streptococcus  pyogenes  hominis  and  the 
Streptococcus  erysipelatis,  and  they  may  therefore  be  regarded  as 
distinct  varieties  (Figs.  89,  90). 

Some  of  the  diseases  and  conditions  in  which  Streptococcus 
pyogenes  has  been  found  may  be  alluded  to  more  in  detail. 

Spreading  Gangrene.— From,  a  case  of  spreading  gangrene,  which  was 
identical  with  Ogston's  erysipelatoid  wound  gangrene,  and  regarded  by 
him  as  the  most  intense  and  dangerous  form  of  erysipelas,  Rosenbach 
obtained  pure-cultivations  of  a  streptococcus  by  incising  the  skin  of  the 
limb,  and  inoculating  tubes  from  the  turbid  reddish  fluid  which  escaped. 
That  the  streptococcus  was  identical  with  Streptococcus  pyogenes  was 
ascertained  by  comparison  with  a  cultivation  derived  from  pus,  of  the 
mode  of  growth,  and  of  the  effect  on  animals. 

Surgical  Fever. — Eiselsberg  proved  the  presence  of  a  streptococcus 
in  the  blood  of  several  cases  of  surgical  fever  in  Billroth's  clinic.  The 
organism  was  identified  by  cultivation  with  Streptococcus  pyogenes. 

Diphtheria. — In  three  cases  of  typical  diphtheria,  Loffler  found  a 
streptococcus.  He  isolated  it  by  cultivation,  found  that  it  was  similar 
in  form,  characters  on  cultivation,  and  effects  after  inoculation,  to 
Fehleisen's  streptococcus  of  erysipelas.  Loffler  was  not  inclined  to 
regard  them  as  identical,  because  Fehleisen  never  found  his  cocci  in  the 
blood-vessels.  Fltigge  named  the  organism  Streptococcus  articulorum, 
and  states,  that  after  subcutaneous  inoculation  or  injection  of  a  cultiva- 
tion in  mice,  a  large  proportion  of  the  animals  die,  and  in  the  sections 
of  the  spleen  and  other  organs  the  streptococci  are  again  seen.  Baum- 
garten  investigated  the  same  subject,  and  decided  that  the  streptococcus 
was  identical  with  Streptococcus  pyogenes. 

Small-pox. — Hlava  has  established  the  presence  of  Streptococcus 
pyogenes  in  the  pustules  of  variola,  and  Gam'  found  streptococci  in  the 
internal  organs  in  a  case  of  variola  haBmorrhagica.  In  a  fatal  case  of 
variola  complicated  with  pemphigus,  Garre  found  a  streptococcus  in 
the  pemphigus  vesicles.  Whether  it  was  identical  with  Streptococcus 
erysipelatis  Garre  left  an  open  question. 

Yellow  Fever. — Babes  observed  the  presence  of  streptococci  in  the 
vessels  of  the  kidney  and  liver  in  yellow  fever.  Cultivation  experiments 
are  wanting.  It  was  probably  a  case  of  secondary  infection  with  Strepto- 
coccus pyogenes. 


SUPPURATION,    PY.EMIA,    SEFHC^MIA,    ERYSIPELAS.  183 

Bilious  Fever, — Babes,  in  a  case  of  fievre  bilieuse  typhoide,  found 
masses  of  streptococci  filling  the  vessels  of  the  liver,  kidney,  and  spleen. 
This  was  probably  another  instance  of  secondary  infection  with  Strepto- 
coccus pyogenes. 

Measles. — From  the  blood  and  inflammatory  post-products  in  measles, 
Babes  isolated  a  streptococcus,  which  he  describes  as  closely  resembling 
the  Streptococcus  pyogenes. 

Ulwiitfrr  l-liijocdi-dit/x. — Wyssokowitsch  found  cocci  in  the  internal 
organs  in  ulcerative  endocarditis,  and  produced  the  disease  in  animals, 
after  injury  to  the  valves,  by  injection  of  Streptococcus  pyogenes  and 
other  organisms.  Weichselbaum,  by  microscopical  research  and  by 
cultivation  experiments,  proved  the  presence  of  Streptococcus  pyogenes 
in  acute  verrucous  endocarditis.  Baumgarten  confirmed  this.  He  found 


FIG.  86.—  ULCERATIVE  ENDOCARDITIS  :  SECTION  OF  CARDIAC  MUSCLE,  x  700  (Kocn). 


Streptococcus  pyogenes  alone  in  one  case  and  accompanied  by  Staphylo- 
coccus  aureus  in  another. 

Broncho-pneumonia, — Thaon  found  a  streptococcus  in  the  lungs  of 
children  in  fatal  cases  of  broncho-pneumonia,  complicating  measles, 
diphtheria,  and  whooping  cough.  It  was  regarded  as  identical  with  the 
streptococcus  isolated  by  Loftier  from  diphtheria.  Frankel  discovered 
a  streptococcus  in  the  lungs  of  a  case  of  true  croup  complicated  with 
broncho-pneumonia,  and  by  cultivation  established  its  identity  with 
Streptococcus  pyogenes. 

Anthrax. — Charrin  found  cocci  in  rabbits,  examined  some  hours  after 
death  from  anthrax.  These,  when  isolated,  produced  death  in  rabbits 
from  septicaemia,  without  suppuration.  Chains  composed  of  from  fifteen 
to  twenty  elements  were  found  in  all  the  organs.  This  was  probably 
another  instance  of  Streptococcus  pyogenes. 

SypJrili*. — Kassowitz  and  Hochsinger  found  the  presence  of  a  strepto- 


184 


INFECTIVE    DISEASES. 


coccus  in  the  tissues  and  internal  organs,  and  especially  in  the  blood- 
vessels, in  fatal  cases  of  congenital  syphilis.  These  observers  regarded 
their  discovery  as  having  an  important  bearing  on  the  etiology  of  syphilis, 
but  Kolisko  pointed  out  that  it  was  only  the  result  of  septic  infection 
with  presence  of  Streptococcus  pyogenes,  as  had  already  been  established 
in  scarlet  fever. 

Cerebro- spinal  Meningitis. — From  the  meningeal  exudation  of  a  case 
of  apparently  idiopathic  cerebro-meningitis,  Banti  found  Streptococcus 
pyogenes  and  Staphylococcus  aureus  and  albus.  The  cocci  probably 
entered  through  an  abscess  of  the  jejunum. 


FlG.    87. — PURE-CULTURES    OF   STREPTOCOCCUS   PvOGENKS. 

a,  On  the  surface  of  nutrient  gelatine  ;  I,  in  the  depth  of  nutrient  gelatine 
c,  on  the  surface  of  nutrient  agar. 


Blepharadenitis  and  Dna-ywyxfix. — Widmark  isolated  by  cultivations 
Streptococcus  pyogenes  and  other  organisms  from  cases  of  blepharadenitis 
and  phlegmonous  dacryocystis.  In  phlegmonous  dacryocystis  Widmark 
found  Streptococcus  pyogenes  almost  exclusively. 

LeukcBmia. — Fliigge  cultivated  a  streptococcus  from  necrotic  patches 
in  the  spleen  of  a  fatal  case  of  leukaemia.  Cultures  corresponded  very 
closely  with  Streptococcus  pyogenes.  Inoculation  in  the  ears  of  rabbits 
produced  similar  results  to  Streptococcus  pyogenes  or  erysipelatis. 
Fliigge  calls  it  Streptococcus  pyogenes  malignus,  but  concludes  that  it 
is  probably  dentical  with  the  streptococcus  from  pus. 


SUPPURATION,    PY^MIA,    SEPTICAEMIA,    ERYSIPELAS.  185 

ERYSIPELAS. 

Erysipelas  is  an  acute  inflammation  of  the  skin,  occurring  in 
connection  with  wounds,  when  it  is  traumatic,  and  on  surfaces 
apparently  sound,  when  it  is  idiopathic,  as  in  erysipelas  of  the 
face.  It  is  highly  contagious  in  surgical  wards,  and  it  gives  rise 
to  rapidly  fatal  puerperal  fever  in  lying-in  hospitals.  In  such  t-ascs 
the  virus  is  obviously  conveyed  from  sick  to  healthy  persons  by 
direct  contact,  or  by  instruments  and  sponges,  or  by  the  hand 
of  the  surgeon,  physician,  or  nurse,  and  possibly  by  the  air. 


FIG.  88.— SECTION  OF  SKIN  IN  ERYSIPELAS. 

v,v,  two  lymphatic  vessels  containing  leucocytes  and  m,in,  streptococci ;  t,  connective 
tissue  ;  a,  connective  tissue  and  wandering  cells,      x  600  (CoRNiL  and  RANVIER). 

Streptococcus  of  Erysipelas. — In  1882,  Fehleiseii  isolated  a 
streptococcus  in  erysipelas,  described  the  appearances  on  cultivation, 
and  maintained  that  it  could  be  distinguished  from  the  streptococcus 
of  suppuration.  Rosenbach  agreed  that  the  two  micro-organisms 
could  be  distinguished  by  parallel  experiments,  and  named  the  one 
Streptococcus  pyogenes  and  the  other  Streptococcus  erysipelatis. 
(Fehleisen).  Rosenbach  asserted  that  the  colonies  of  the  latter 
were  more  opaque  and  whiter  than  those  of  Streptococcus  pyogenes 
and  the  growth  more  marked  in  the  depth  of  nutrient  gelatine, 
while  microscopically  the  chains  were  better  marked  and  larger,  and 
the  individual  cocci  larger  than  in  Streptococcus  pyogenes.  Others 
who  investigated  this  subject  could  not  distinguish  them  with 
certainty,  either  by  their  morphological  or  cultural  characters  or 
effects  on  inoculation.  Passet  found  that  inoculation  of  Strepto- 
coccus pyogenes  induced  a  condition  very  similar  to  that  produced  by 
inoculation  of.  Streptococcus  erysipelatis.  Hoffa  arid  Hajek  described 
minute  differences,  but  Biondi  and  Eiselsberg  failed  to  confirm  these. 


186  INFECTIVE   DISEASES. 

Baumgarten  failed  to  prove  any  essential  difference.  Mitchell 
Prudden  found  that  Streptococcus  pyogenes  injected  into  the  sub- 
cutaneous .  tissue  of  the  ears  of  rabbits,  produced  in  one  no  effect  ; 
in  four,  slight  transient  redness  ;  in  five,  local  redness  followed  by 
abscess ;  in  twelve,  well-marked  erysipelatous  redness,  followed  by 
complete  resolution  in  seven,  abscess  in  three,  and  death  in  two. 
Passet,  Biondi,  Eiselsberg,  Baumgarten,  and  Mitchell  Prudden 
concluded  that,  in  their  morphological,  biological,  and  pathogenic 
characters,  so  far  as  animals  are  concerned,  the  two  organisms  are 
practically  identical. 

The  author  investigated  the  morphology  and  cultural  characters 
of  the  Streptococcus  erysipelatis,  which  he  had  isolated  from  a 
typical  case.  This  result  cleared  up  the  conflicting  statements 
which  had  been  made  by  different  observers.  By  carrying  out 
absolutely  parallel  experiments,  the  Streptococcus  pyogenes  and 
Streptococcus  erysipelatis  were  unquestionably  distinguishable,  as 
Fehleisen  and  Rosenbach  had  asserted.  In  both  cases,  however, 
inoculation  of  a  trace  of  a  culture  from  a  solid  medium  produced 
only  transient  redness.  Injection  hypodermically  of  a  broth-culture 
produced  in  both  cases  a  spreading  erysipelatous  redness,  followed 
by  suppuration.  It  was  found  that  primary  cultures  of  the  two 
micro-organisms,  cultivated  under  precisely  the  same  conditions, 
differed  in  the  size  and  character  of  their  chains,  in  the  size  of  the 
individual  elements,  in  the  greater  opacity  of  the  colonies  of  Strepto- 
coccus erysipelatis,  in  a  greater  tendency  to  confluence,  and  in 
a  more  rapid  growth.  The  author  found  that  the  difference  was 
most  marked  in  broth-cultures.  Abundant  flocculi  were  formed  by 
Streptococcus  pyogenes  ;  a  powdery  deposit  with  special  tendency  to 
form  a  granular  adhesive  film  at  the  bottom  of  the  culture  flask, 
in  the  case  of  the  streptococcus  of  erysipelas.  Lastly,  they  differed 
in  their  power  of  resisting  germicides. 

Fehleisen  inoculated  patients  in  hospital  suffering  with  malignant 
growths,  and  produced  a  typical  erysipelas  with  sub-cultures  after 
an  incubation  of  from  sixteen  to  twenty  hours.  The  disease  was 
marked  by  rigors,  fever,  and  general  disturbance.  Patients  who  had 
recently  suffered  from  erysipelas  had  an  immunity. 

Emmerich  succeeded  in  proving  the  presence  of  streptococci  in 
the  air  of  a  hospital  where  erysipelas  had  broken  out.  These  cocci 
in  their  form,  their  characters  on  cultivation,  and  their  inoculation 
results,  were  identified  with  the  Streptococcus  erysipelatis.  It  is  riot 
therefore  exclusively  parasitic. 

Streptococci  identical  or  agreeing  very  closely  in  their  description 


SUPPURATION,    PY^MIA,    SEPTICAEMIA,   ERYSIPELAS.  187 

\vitli  Streptococcus  pyogenes.  have  been  found  in  cattle  plague,  foot 
and  mouth  disease,  strangles,  contagious  mammitis  in  cows,  and 
progressive  tissue  necrosis  in  mice,  and  they  will  be  referred  to  fully 

in  subsequent  chapters. 

EXAMINATION  AND  CULTIVATION  OF  STREPTOCOCCI. 
Cover-glass  preparations  can  be  stained  with  the  watery  solutions 
of  the  aniline  dyes.     In  some  cases  very  beautiful  preparations  can 
be  obtained  by  using  Neelsen's    solution,  and  removing   excess   of 


b.  c.  .  d. 

FIG.  89. — STREPTOCOCCUS  PYOGENES  HOMINIS.     Pure-cultures  on  nutrient 


gelatine. 


«,  Sub-culture  from  agar. 
c,  Sub-culture  from  milk. 


b,  Sub-culture  from  broth. 
d,  Sub-culture  from  milk. 


stain  by  rinsing  in  alcohol.  To  examine  pus,  milk,  or  broth,  take 
nn  ordinary  platinum  needle  bent  at  the  extremity  into  a  booklet. 
Dip  it  into  the  liquid  to  be  examined,  and  spread  it  on  a  cover- 
idass  into  as  thin  a  film  as  possible;  the  preparation  is  treated 
in  the  ordinary  way,  that  is 'to  say,  the  film  is  allowed  to  dry, 
and  the  cover  is  taken  up  with  forceps,  and  passed  three  tinio 
through  the  flame  with  its  prepared  side  uppermost. 

Gram's  Method  with  Eosin. — In  this  way  the  streptococci  art- 
stained  blue,  and  stand  out  in  marked  contrast  to  the  rest  of  the 
preparation.  Use  freshly  prepared  solution.  Float  tbe  cover-glasses 


188 


INFECTIVE    DISEASES. 


on  the  solution  for  ten  minutes  to  half  an  hour,  then  trai.sfer  them 
to  iodine-potassic-iodide  solution,  until  they  assume  the  colour  of  a 
tea  leaf ;  then  immerse  them  in  alcohol  until  they  are  decolorised  ; 
dip  them  in  an  alcoholic  solution  of  eosin  for  a  few  moments,  and 
then  transfer  them  to  clove  oil  to  clarify  the  film  ;  to  remove  the 
clove  oil  gently  press  the  cover  between  two  layers  of  clean  filter 
paper,  then  mount  in  xylol  balsam. 

A  good  method  for  cultivating  streptococci  is  to  employ  a  steril- 
ised looped  platinum  wire,  and  to  spread  a  droplet,  for  example,  of 
pus  or  blood,  over  the  surface  of  nutrient  agar-agar  solidified  obliquely. 


a-  '>•  c. 

FIG.  90.— STREPTOCOCCUS  PYOGENES  Bovis.     Pure-cultures  on  nutrient 

gelatine. 

a,  Sub-culture  from  agar.  b,  Sub-culture  from  broth. 

c,  Sub-culture  from  milk.  d,  Sub-culture  from  milk. 

The  tubes  are  then  placed  in  the  incubator  at  37°  C.  ;  the  strepto- 
cocci will  appear  in  the  course  of  two  or  three  days  in  the  form 
of  minute  dotted  colonies.  If  present  alone,  and  in  considerable 
quantities,  the  inoculated  surface  will  exhibit  a  pure  cultivation 
consisting  of  a  number  of  such  colonies,  whilst  a  flocculent  mass  is 
observed  in  the  liquid  which  collects  at  the  bottom  of  agar-agar 
tubes ;  this  flocculent  mass  will  be  found  to  be  composed  of  chains. 
From  such  a  tube  inoculate  a  number  of  the  small  flasks  employed 
in  Pasteur's  laboratory  for  cultivations  in  liquids.  In  this  way 
a  number  of  pure- cultivations  in  milk  and  broth  are  established, 
which  can  le  readily  examined  from  time  to  time.  From  a  pure- 


SUPPURATION,    PYAEMIA,    SEPTICAEMIA,    ERYSIPELAS.  189 

cultivation  in  broth  or  agar-agar  tubes  of  nutrient  gelatine  can 
be  inoculated.  Cover-glass-preparations  from  the  growths  on  solid 
media  can  be  made  in  the  usual  way,  and  stained  with  either  a 
watery  solution  of  fuchsine  or  gentian  violet  :  but  to  stain  prepa- 
rations made  from  milk  or  broth,  or  from  the  liquid  in  agar-agar 
tubes,  use  the  method  of  Gram  ;  the  stain  will  then  be  removed, 
except  from  the  streptococci,  .and  very  beautiful  preparations 
result. 

GONORRHCEA. 

Gonorrhoea  is  the  result  of  a  catarrhal  inflammation  of  the 
mucous  membrane  of  the  urethra,  vagina,  or  conjunctiva  caused  by 
a  characteristic  pyogenic  organism  discovered  by  Neisser  in  1879. 

Gonococcus  of  Neisser.  —  Cocci,  usually  in  pairs  1-6  p  in 
length,  -8  ^  in  width,  and  tetrads,  with  those  surfaces  of  the  com- 
ponent elements  which  are  in  contact,  flattened.  The  elements 
are  more  or  less  kidney-shaped,  and  are  separated  by  a  clear 
unstained  interval.  They  are  found  free  in  the  pus  and  also  in 
the  interior  of  the  pus  cells.  They  stain  with  the  aniline  dyes, 
but  are  decolorised  by  Gram's  solution.  They  do  not  grow  on 
the  ordinary  media,  such  as  gelatine,  agar,  and  potato,  in  marked 
contrast  to  the  common  pyogenic  cocci  ;  but  Bumm  succeeded  in 
obtaining  a  cultivation  by  using  human  blood  serum,  which  was 
procured  for  the  purpose  from  the  placenta.  They  give  rise  to 
a  very  delicate  growth  in  the  form  of  an  almost  invisible  film, 
with  a  moist  appearance,  which  attains  its  full  development  in 
a  few  days.  Steinschneider  used  human  blood  serum  and  aufar 
incubated  at  35°  0. 

Krall  recommended  either  agar  with  grape-sugar  and  blood  serum, 
or  the  same  mixture  with  the  addition  of  5  per  cent,  glycerine. 
Others  have  employed  nutrient  agar  with  the  surface  moistened  with 
sterilised  human  blood.  More  recently  Keifer  has  been  successful 
with  a  medium  which  is  prepared  in  the  following  way  :  a  si-it  ic- 
fluid  is  filtered  arid  sterilised  by  Tyndall's  process,  to  this  is  add«'d 
an  equal  quantity  of  the  following  mixture,  agar  3'5,  peptone  5, 
glycerine  2,  salt  '5  (per  cent.).  The  ascitic  agar  is  solidified  in  a 
Petri's  dish,  and  the  culture  incubated  at  36°  C. 

They  have  also  been  cultivated  in  albumin  from  plovers'  eggs, 
and  in  the  fluid  obtained  from  a  case  of  synovitis  of  the  knee  joint. 

Inoculation  of  rabbits,  dogs,  horses,  and  monkeys,  has  been 
invariably  unsuccessful,  but  sub-cult  -nros  produce  the  disease  in 
the  health  urethra. 


OF    TBK 

UNIVERSITY 


190  INFECTIVE    DISEASES. 

The  cocci  are  found  in  pus  from  the  urethra  and  other  mucous 
membranes  affected  by  the  disease.  They  have  also  been  found  in 
urethral  and  inguinal  abscesses  in  association  with  Staphylococcus 
pyogenes  aureus. 

METHOD  OF  STAINING. 

Cover-glass  preparations  are  made  in  the  usual  way,  and 
double  stained  with  Loffler's  methylene  blue,  and  eosiii. 

Schiitz  recommends  floating  the  cover-glasses  for  five  or  ten 
minutes  in  a  saturated  solution  of  methylene  blue  in  5  per  cent, 
solution  of  carbolic  acid.  They  are  washed  in  water,  rinsed  in  very 
weak  acetic  acid,  and  again  washed  in  water.  Safranin  may  be 
used  as  a  contrast  stain. 


FIG.  91.— GONOCOCCUS  x  800  (BUMM). 
a,  free  cocci ;  b,  cocci  in  pus  cells  ;  c,  epithelial  cell  containing  cocci. 

EGYPTIAN  OPHTHALMIA. 

There  are  two  forms  of  ophthalmia  in  Egypt,  one  associated 
with  Gonococcus  and  the  other  with  a  bacillus  closely  resembling 
the  bacillus  of  mouse-septicaemia,  but  there  are  minute  differences. 

Bacillus  of  Ophthalmia  (Koch  and  Kartulis).  Minute  rods 
which  do  not  grow  on  gelatine  but  readily  on  blood  serum  and 
nutrient  agar,  forming  a  plainly  visible,  whitish-grey  shining  growth. 
Animals  are  insusceptible,  but  cultures  produced  the  disease  in  the 
human  conjunctiva  in  two  out  of  six  cases. 


CHAPTER   XIV. 

ANTHRAX. 

ANTHRAX  is  a  very  fatal  malady,  and  most  irregular  in  its  be- 
haviour. At  one  time  it  attacks  only  one  or  two  animals,  and 
at  another  time  it  will  destroy  nearly  all  the  stock  on  a  farm. 
Farmers  formerly  regarded  the  disease  as  non- communicable,  and 
possibly  the  result  of  excessive  or  improper  feeding,  or  faulty  sani- 
tation, or  of  climatic  conditions  over  which  no  control  could  be 
exercised.  It  is  obvious  that  so  long  as  the  disease  was  regarded  as 
the  result  of  unknown  conditions,  no  explanation  could  be  given  of 
its  recurrence  from  time  to  time,  or  of  certain  animals  contracting 
the  disease  and  others  not,  and  no  measures  of  any  use  could  be 
suggested  to  cope  with  an  outbreak. 

Anthrax  has  always  been  more  prevalent  on  the  Continent  than 
in  England,  and  this  to  some  extent  accounts  for  the  fact  that  it  has 
received  greater  attention  abroad.  In  France,  Germany,  Hungary, 
Russia,  and  in  India  and  Persia,  anthrax  at  times  produces  wide- 
spread losses.  In  Siberia  it  is  still  known  on  this  account  as  the 
Siberian  Plague. 

On  the  Continent  there  are  certain  localities  known  as  anthrax 
districts  on  account  of  their  reputation  for  anthrax — for  example,  in 
the  Upper  Bavarian  Alps  in  Germany  and  in  Auvergne  in  France. 

In  1849,  Pollender  happened  to  examine  the  blood  of  a  cow 
after  death  from  anthrax,  and  discovered  peculiar  rod-like  bodies 
among  the  blood  cells.  The  same  observation  was  made  independ- 
ently by  Brauell  and  Davairie  about  the  same  time,  but  the  greatest 
importance  must  be  attached  to  the  publication  of  Davaine's  further 
researches  in  1863.  Many  ridiculed  the  discovery  of  bacilli,  and 
stoutly  maintained  that  they  were  only  blood  crystals  or  accidental 
structures  of  no  importance. 

For  many  years  very  little  progress  was  made,  and  the  statements 
of  other  observers  who  were  able  to  verify  and  add  to  Pollender's 
and  Davaine's  discoveries,  were  still  received  with  scepticism. 

191 


192 


INFECTIVE    DISEASES. 


Within  the  last  few  years  a  great  change  of  opinion  has  taken 
place.  Bacteriologists  have  investigated  the  whole  subject,  so  that 
at  the  present  day  we  know  exactly  the  cause  of  anthrax. 

Bacillus  anthracis  (Bacteridie  du  charbon,  Bacillus  of  splenic 
fever,  Wool-sorters'  disease,  or  malignant  pustule). — Rods  5  to  20  ^ 
long  and  1  to  1'25  p  broad,  and  threads;  spore-formation  present. 
As  a  thorough  knowledge  of  the  life-history  of  this  bacillus  is  of  the 
greatest  importance,  the  various  steps  to  be  followed  in  a  practical 
study  of  it  will  be  successively  treated  in  detail.  Its  morphological 


*  vf.m 

™  /a£8 


FIG.  92. — BACILLUS  AXTHRACIS,  x  1200.     Blood  corpuscles  and  bacilli 
unstained ;  from  an  inoculated  mouse  (FRANKEL  and  PFEIFFER). 

and  biological  characteristics  have  been  very  completely  worked  out, 
and  it  serves  as  an  excellent  subject  for  gaining  an  acquaintance 
with  most  of  the  methods  employed  in  studying  micro-organisms. 

A  mouse  inoculated  with  the  bacillus  or  its  spores  will  die  in 
from  twenty-four  to  forty-eight  hours,  or  more  rarely  in  from 
forty-eight  to  about  sixty  hours. 

Examination  after  Death. — The  spleen  is  found  to  be  considerably 
enlarged,  and  may  be  removed,  and  examined  by  making  cover-glass 
preparations,  inoculations  in  nutrient  media,  and  subsequently 
sections. 

Cover-glass  Preparations. — In  cover-glass  preparations  of  the 
blood  of  the  spleen  the  bacilli  are  found  in  enormous  numbers. 
Preparations  should  also  be  made  with  blood  from  the  heart  and 
with  the  exudation  from  the  lungs  and  other  organs  ;  it  will  be 


DESCRIPTION   OF   PLATE   Y. 
Bacillus  Anthracis. 

FIG.  1. — From  a  cover-glass  preparation  of  blood  from  the  spleen  of  a  guinea- 
pig  inoculated  with  blood  from  a  sow.  x  1200.  Powell  and  Lealand's 
apochromatic  T\  Horn.  imm.  E.  P.  10. 

FIG.  2. — From  a  section  of  a  kidney  of  a  mouse.  Under  a  low  power  the 
preparation  has  exactly  the  appearance  of  an  injected  specimen.  Under 
higher  amplification  the  bacilli  are  seen  to  have  threaded  their  way  along 
the  capillaries  between  the  tubules,  and  to  have  collected  in  masses  in 
the  glomeruli.  Stained  with  Gram's  method  (gentian-violet),  and  eosin. 
x  500. 

FIG.  3. — Bacillus  anthracis  and  Micrococcus  tetragenus.  From  a  section  from 
the  lungs  of  a  mouse  which  had  been  inoculated  with  anthrax  three  days 
after  inoculation  with  Micrococcus  tetragenus.  A  double  or  mixed  infection 
resulted.  Anthrax-bacilli  occurred  in  vast  numbers,  completely  filling  the 
small  vessels  and  capillaries,  and  in  addition  there  were  great  numbers 
of  tetrads.  Stained  by  Gram's  method  (gentian-violet),  and  with  eosin. 
x  500. 


Plate  V. 


-^  \ 


y/s  js»  * 
I'xC^  / 


Fig 


Fig 


2. 


Fig  3. 


BACILLUS    ANTHRACIS 


ANTHRAX. 


193 


noted  that  in  these  the  bacilli  are  present  in  very  small  numbers,  or 
altogether  absent.  The  bacilli  should  be  examined  both  unstained 
and  stained.  The  rods  are  straight  or  sometimes  curved ;  rigid  and 
motionless.  They  can  be  stained  with  a  watery  solution  of  any  of 
the  aniline  dyes,  and  are  then  seen  to  be  composed  of  segments  with 
their  extremities  truncated  at  right  angles ;  between  the  segments 
a  clear  linear  space  exists,  which  gives  them  a  characteristic  appear- 
ance (Plate  V.,  Fig.  1).  By  double  staining,  with  Gram's  method  and 
eosin,  the  rods  are  seen  to  consist  of  a  membrane  or  hyaline  sheath 
with  protoplasmic  contents. 

Drop-cultures. — A  little  of  the  blood  from  the  spleen  or  heart 
may  be  employed  to  inoculate  sterilised  broth  or  blood  serunu 
Several  of  these  cultures  should  be  prepared,  and  some  of  them 
placed  in  the  incubator,  and  examined  at  intervals  of  a  few  hours. 
It  will  be  observed  that  the  rods  grow  into  long  homogeneous  fila- 
ments, which  are  twisted  up  in  strands,  and  partly  untwisted  in  long 
and  graceful  curves.  The  filaments  begin  to  swell, 
become  faintly  granular,  and  bright,  oval  spores 
develop  (Plate  1).  The  cultures  in  the  incubator 
develop  rapidly.  A  temperature  of  30°  to  37°  C. 
is  the  most  favourable  for  spore-formation.  The 
spores  are  eventually  set  free,  and  by  making 
a  fresh  cultivation,  or  by  injecting  them  into  a 
mouse  or  guinea-pig,  they  germinate  again  into 
the  characteristic  bacilli,  which  in  their  turn 
grow  into  filaments  and  spores.  When  the  spore 
germinates  it  swells,  the  envelope  becomes  jelly- 
like,  and  gives  way  at  one  or  other  pole,  and  the 
contents  escape  and  grow  into  a  rod. 

Test-tube  Cultivations  in   Nutrient    Gelatine. — 
Typically  characteristic  appearances  are  obtained 
by  inoculating  a  5  to  8  per  cent,  nutrient  gelatine. 
A  whitish  line  develops  in  the  track  of  the  inocu- 
lating needle,  and  from  it  fine  filaments  spread  out 
in  the  surrounding  medium  (Fig.  93).     The   fila-  FIG.  93.— PURE  CUL- 
ments  are  more  easily  observed  with  a  magnifying      TIVATION   OF  BA- 
glass.     In  a  more  solid  nutrient  gelatine  the  growth 
appears  only  as  a  thick  white  thread.     As  lique-      LATINE. 
faction  of  the  gelatine  progresses,  these  appearances 
gradually   alter,    and   the   growth  subsides   to   the   bottom  of  the 
tube  as  a  white  flocculent  mass.     In  exhausted  culture- media,  and 
sometimes  in  the  blood,  filaments  are  seen  in  a  state  of  degeneration. 

13 


194 


INFECTIVE    DISEASES. 


This  has  also  been  observed  in  sections  of  the  internal  organs  of  a 
rabbit  which  had  been  inoculated  with  the  anthrax  bacillus  and  had. 
died  of  septicaemia  the  following  morning. 

Test-tube  Cultivations  in  Nutrient  Agar-agar. — Cultivated  upon  a 


FIG.  94.— COLONIES  OF  BACILLUS  ANTHRACIS,  x  80  (FLUGGE). 
a,  after  24  hours ;  b,  after  48  hours. 

sloping  surface  of  nutrient  agar-agar  a  viscous  snow-white  layer  is 
developed,  but  without  access  of  air  no  cultivation  can  be  obtained, 
the  bacilli  being  aerobic.  This  can  be  demonstrated  by  completely 

embedding  a  piece  of  lung  or  spleen 
pulp  containing  bacilli,  in  nutrient 
agar-agar  (p.  22). 

Potato  -  cultivations.  —  In  about 
thirty -six  to  forty-eight  hours  a  creamy- 
white  or  very  faintly  yellowish  layer 
forms  over  the  inoculated  surface, 
usually  with  a  translucent  edge,  and 
sometimes  a  strong,  penetrating  odour 
of  sour  milk. 

Plate-cultivations. — From  the  spleen 
or  blood  of  the  heart  cultivations  may 
be  made  in  nutrient  gelatine  on  plates. 
The  colonies  develop  in  about  two  days,  according  to  the  temperature 
of  the  room.  They  appear  to  the  naked  eye  as  little  white  spots 


FIG.    95.— IMPRESSION-PREPARA 
TION  OF  A  COLONY,   X  70. 


ANTHRAX. 


195 


or  specks,  which,  on  examination  with  a  low  power  of  the  microscope 
and  small  diaphragm,  exhibit  two  distinct  forms.  One  form,  on 
careful  focussing,  has  the  appearance  of  a  little  compact  ball  of 


FIG.  96.— MARGIN  OP  A  COLONY,  x  250 

twisted  threads ;  in  the  other,  liquefaction  of  the  gelatine  has 
commenced,  and  the  threads  spread  out  like  locks  or  plaits  of 
hair  in  the  neighbouring  gelatine.  These  appearances  are  perfectly 
characteristic  (Figs.  94,  96). 

Cover-glass  Impressions. — The  plate- cultivations  should  be  also 
examined  as  soon  as  the  colonies 
appear,  by  making  cover-glass  im- 
pressions (Fig.  95).  The  filaments, 
examined  with  a  high  power,  will  be 
seen  to  consist  of  a  number  of  rods 
or  segments  which  are  perfectly 
regular  in  form.  On  the  other 
hand,  filaments  from  a  tube-cultiva- 
tion in  a  solid  medium  will  often  be 
found  to  be  composed,  not  only  of 
rods,  but  here  and  there  of  the  so- 
called  involution-forms  (Fig.  97).  FIG.  97.— FILAMENTS  WITH  OVAL  AND 

„  ,     ,  .  i  IRREGULAR  ELEMENTS,     X    800. 

From    cultures     m    gelatine     and 

glycerine  agar,  very  striking  preparations  are  sometimes  'obtained, 

with  numerous  large  spherical   and  lemon-shaped  elements.     In   a 


196  INFECTIVE    DISEASES. 

cover-glass  preparation  from  a  potato-culture  the  individual  segments 
will  be  found  to  have  a  great  tendency  to  be  isolated  one  from  the 
other,  and  there  is  copious  spore-formation. 

Preservation  of  Spores. — .Spores  may  be  preserved  simply  by  allow- 
ing anthrax  blood  to  dry  and  then  sealing  it  in  a  tube.  The  spores 
from  a  potato-cultivation  are  treated  as  follows  : — The  inoculated 
surface  bearing  the  creamy  cultivation  is  sliced  off  in  a  thin 
layer,  and  is  mashed  up  with  distilled  water  in  a  glass  capsule. 
Sterilised  silk-thread  is  cut  up  into  lengths  of  about  a  quarter  of 
an  inch,  and  allowed  to  soak  in  the  paste  for  some  hours,  under  a 
bell-glass.  The  threads  are  then  picked  out  with  a  pair  of  forceps, 
and  laid  upon  a  sterilised  glass  plate,  covered  with  a  bell-glass, 
and  allowed  to  dry.  From  the  plate,  when  perfectly  dry,  they 
are  transferred  to  a  small  test-tube,  which  can  be  plugged  with 
cotton-wool,  or  sealed  in  the  Bunsen  burner. 

Examination  of  the  Tissues. — The  organs  should  be  hardened 
in  absolute  alcohol,  and  sections  prepared  and  stained  by  the 
ordinary  methods.  The  method  of  Gram  is  the  most  instructive, 
and  eosin  a  very  satisfactory  contrast  stain.  The  capillaries  in 
the  lungs,  liver,  kidney,  spleen,  skin,  mucous  membrane,  etc.,  will 
be  found  to  contain  bacilli.  In  some  cases  the  bacilli  are  so 
numerous  that  a  section  under  a  low  power  has  the  appearance 
of  an  injected  specimen. 

Inoculation  of  Animals. — A  thread  containing  spores,  a  drop  of 
blood  from  an  infected  animal,  or  a  minute  portion  of  a  cultivation, 
introduced  under  the  skin  of  a  mouse  or  guinea-pig,  causes  a  fatal 
result,  as  a  rule,  in  from  twenty-four  to  forty-eight  hours.  Sheep 
fed  upon  potatoes  which  have  been  the  medium  for  cultivating  the 
bacillus,  die  in  a  few  clays.  Goats,  hedgehogs,  sparrows,  cows,  horses, 
swine,  and  dogs  are  all  susceptible.  Rats  are  infected  with  diffi- 
culty. Frogs  and  fish  have  been  rendered  susceptible  by  raising  the 
temperature  of  the  water  in  which  they  lived.  Cats,  white  rats, 
and  Algerian  sheep  have  an  immunity  from  the  disease. 

Attenuation  of  the  Virus.  —  Toussaint  attenuated  cultures  by 
exposing  them  for  ten  minutes  to  55°  0.  Pasteur  obtained  a  similar- 
result  by  resorting  to  lower  degrees  of  temperature ;  and  Koch, 
Gaffky,  and  Loffler  concluded  from  their  experiments,  that  from 
42°  to  43°  C.  the  bacillus  was  most  easily  deprived  of  its  poisonous 
properties.  By  cultivating  the  bacillus  in  neutralised  broth  at 
42°  to  43°  C.  for  about  twenty  days,  the  infecting  power  is  weakened, 
and  animals  inoculated  with  it  (premier  vaccin)  are  protected  against 
the  disease.  To  obtain  a  still  more  perfect  immunity,  they  are 


ANTHRAX.  197 

inoculated  a  second  time  with  material  (deuxieme  vaccin)  which  has 
been  less  weakened.  The  animals  are  then  protected  against  the 
most  virulent  anthrax,  but  only  for  a  time.  From  a  weakened 
culture,  according  to  Klein,  new  cultures  of  virulent  bacilli  can  be 
started,  and  a  culture  that  can  be  used  as  a  vaccine  for  sheep  kills  a 
guinea-pig,  and  then  yields  bacilli  that  are  fatal  to  sheep. 

The  virulence  of  the  bacillus  is  also  altered  by  passing  the 
bacillus  through  different  species  of  animals.  The  bacillus  of  sheep 
or  cattle  is  fatal  when  re-inoculated  into  sheep  or  cattle;  but  if 
inoculated  in  mice,  the  bacilli  then  obtained  lose  their  virulence 
for  sheep  or  cattle,  only  a  transitory  illness  results,  and  the  animals 
are  protected  for  a  time  against  virulent  anthrax. 

Exposure  to  a  temperature  of  55°  C.,  or  treatment  with  *5  to 
1  per  cent,  carbolic  acid,  deprives  the  bacilli  of  their  virulence. 

Chauveau  obtained  a  similar  result  by  cultivating  the  bacillus  at 
38°  or  39°  C.  under  a  pressure  of  eight  atmospheres.  The  possibility 
of  mitigating  the  virus  depends  upon  the  species  of  animal ;  rodents 
cannot  be  rendered  immune  by  any  known  anthrax  vaccine.  The 
nature  of  the  toxic  products  has  been  described  in  a  previous  chapter 
(p.  42). 

METHODS  OF  STAINING  THE  BACILLUS  ANTHRACIS. 

Cover-glass  preparations  of  blood,  etc.,  can  be  stained  with  a 
watery  solution  of  any  of  the  aniline  dyes,  or  with  Neelsen's  solution 
and  subsequent  treatment  with  alcohol  (p.  87).  The  preparations 
may  be  dried  and  mounted  permanently  in  Canada  balsam,  but  the 
typical  appearances  are  best  observed  in  freshly  stained  specimens 
examined  in  water. 

The  sheath  and  protoplasmic  contents  can  be  demonstrated  in 
cover-glass   preparations    from   the 
blood   or   spleen  which    have   been 
stained  with  eosin  after  the  method 
of  Gram. 

Spores  must  be  stained  by  the  ^ 

special  methods    already  described.          /^^        C^ 
The  most  satisfactory  preparations 
are  obtained  by  double-staining  with         FIG.  98.— SPORES  OF  BACILLUS 
Ziehl-Neelsen  solution  and  methy-  ANTHRACIS     STAINED     WITH 

,  ,       /T^.      ~v  GENTIAN  VIOLET,    x   1500. 

lene  blue  (Fig.  / ). 

Tissue  sections  are  best  stained  by  the  method  of  Gram,  and 
after-stained  with  eosin,  picrocarminate  of  ammonia,  or  picro-lithium- 
carmine. 


198  INFECTIVE    DISEASES. 


ORIGIN  AND  MODE  OF  SPREAD. 

As  every  outbreak  of  anthrax  is  the  result  of  the  introduction 
into  the  system  of  the  bacilli,  the  question  naturally  arises,  how 
are  they  introduced  on  the  farm  ?  Where  do  they  come  from  ?  and 
what  are  the  channels  of  infection  ? 

The  spores  of  the  bacilli  may  get  into  the  soil,  and  may  remain 
there  in  a  dormant  state  for  many  years.  The  spores  were  believed 
by  Pasteur  to  be  taken  up  by  earth-worms,  carried  to  the  surface 
and  deposited  in  their  castings.  Animals  grazing  are  thus  liable 
to  be  infected;  but  Koch's  experiments  tended  to  disprove  this 
theory.  Anthrax  has  been  known  to  break  out  among  cattle 
grazing  on  a  field  where  several  years  previously  some  Russian 
hides  from  infected  animals  had  been  buried.  By  some  means  or 
other  the  spores  may  contaminate  the  grass,  and  hay  imported 
from  an  anthrax  district  may  start  the  disease  on  a  farm  on  which 
it  had  never  been  known  to  occur.  The  spores  may  in  a  similar 
way  be  introduced  with  blood  manure  and  bone  manure,  and  with 
refuse  used  as  manure.  The  skin,  hair,  wool,  hoofs,  and  horns  of 
infected  animals,  if  soiled  with  blood,  are  contaminated  by  the 
bacillus. 

Another  way  in  which  the  disease  can  be  communicated  may 
be  illustrated  by  the  transmission  of  the  disease  to  man.  Those 
who  handle  carcasses,  wool  or  hides  of  infected  animals  are  liable  to 
contract  the  disease.  Slight  scratches,  cuts,  bites,  and  pimples,  may 
readily  be  inoculated  with  the  bacilli  or  their  spores.  Veterinary 
surgeons,  butchers,  herdsmen,  cattle  drovers — in  fact,  all  those  whose 
occupation  leads  them  to  cut  open  or  skin  cattle,  sheep,  or  horses,  or 
to  handle  hides  and  wool — are  liable  to  fall  victims  to  this  disease. 

In  one  case  which  was  brought  to  the  author's  notice,  a  veteri- 
nary surgeon  had  been  called  to  see  a  bullock  which  had  died 
suddenly  in  a  meadow.  A  post-mortem  examination  was  made,  and 
the  veterinary  surgeon  wiped  his  hands,  which  were  soiled  with 
blood,  on  some  rough  grass,  and  then  washed  them  in  a  stream. 
The  sedgy  grass  made  some  small  cuts  on  his  fingers,  and  the 
result  was  that  he  was  simultaneously  inoculated  with  the  blood 
of  the  bullock.  Local  anthrax  followed,  two  of  his  fingers  were 
amputated,  and  he  fortunately  recovered.  In  another  case  a  butcher 
dressed  the  carcass  of  a  beast  which  had  died  suddenly,  and  while 
doing  so  scratched  a  pimple  on  his  neck.  An  anthrax  pustule 
developed,  and  after  a  very  serious  illness  he  also  recovered ;  but 
in  many  cases  the  attack  is  fatal.  "  Wool  -sorters'  disease"  is 


ANTHRAX.  199 

anthrax  of  the  lungs.  Bales  of  foreign  wool  contain  not  only  wool 
from  living  sheep,  but  wool  which  has  been  clipped  from  skins  of 
dead  sheep.  If  any  of  the  sheep  died  from  anthrax  the  wool  is 
sure  to  be  contaminated  with  blood  containing  the  bacilli,  and  then 
wool-sorters  engaged  in  picking  the  wool  readily  inoculate  them- 
selves through  a  scratch  or  pimple,  or  by  inhaling  the  spores.  In 
many  cases  Wool-sorters'  disease  is  fatal. 

A  farm  may  become  extensively  infected  by  the  living  animal. 
Blood  containing  the  bacilli  may  be  discharged  from  the  mouth  and 
nostrils,  or  be  passed  with  the  contents  of  the  intestinal  canal 
and  bladder.  The  droppings  contaminate  the  pasture  or  byre,  and 
spore  formation,  especially  in  warm  weather,  quickly  takes  place. 
From  this  cause  the  disease  may  not  only  be  conveyed  to  healthy 
cattle  grazing  with  infected  animals,  but  fresh  cases  may  occur,  year 
after  year,  on  the  same  far  in,  and  if  hay  is  cut  and  sold  off  the  farm, 
other  cattle  at  a  distance  are  similarly  infected.  If  the  flooring  of 
cattle  sheds  is  once  soiled  by  infected  animals  it  is  easy  to  account 
for  those  otherwise  mysterious  outbreaks  which  occur  when  the 
cattle  are  taken  in  for  the  winter. 

Another  source  of  danger  arises  when  blood  from  a  diseased 
animal  is  washed  into  brooks  or  streams,  for  thus  the  disease  may 
be  carried  to  farms  in  which  it  was  previously  unknown. 

PREVENTIVE  MEASURES. 

Early  recognition  and  prompt  action  are  essential  to  prevent  the 
spread  of  any  communicable  disease. 

Unfortunately  in  the  case  of  anthrax  only  too  often  the  very 
first  indication  of  the  existence  of  the  disease  is  the  sudden  death 
in  the  pasture  or  byre  of  an  apparently  healthy  beast,  or  possibly 
of  one  or  more  sheep.  Nevertheless,  the  importance  of  being  able 
to  recognise  any  early  indications  is  very  great,  because  an  im- 
mediate and  careful  examination  should  at  once  be  made  of  the 
stock  on  the  farm,  and  suspicious  cases  isolated  from  the  rest.  The 
stock-man  may  notice  that  one  or  two  animals  tend  to  keep  away 
from  the  others.  They  look  dull  and  cease  feeding,  and  possibly 
shivering  may  be  observed.  In  horses  swelling  of  the  throat  may 
occur,  and  in  some  places,  there  is  discharge  of  blood  from  the 
orifices.  Death  follows  the  appearance  of  these  symptoms  in  a 
few  hours,  and  often  with  startling  suddenness.  Cattle  die  rapidly, 
but  sheep,  though  rapidly  contracting  the  disease,  do  not  as  a  rule 
die  so  suddenly. 


200  INFECTIVE    DISEASES. 

The  characteristic  sign  after  death  is  enlargement  of  .the  spleen 
to  three  or  four  times  its  natural  size.  It  is  not  only  -enlarged, 
but  extremely  soft  and  dark  in  colour.  Blood  spots  are  visible  on 
the  internal  organs  generally,  and  the  intestine  often  contains  a 
quantity  of  blood.  The  examination  of  a  drop  of  blood  will  show 
under  the  microscope  the  characteristic  bacilli.  It  is,  however, 
quite  unnecessary  to  make  an  elaborate  post-mortem  examination 
in  order  to  satisfy  oneself  whether  the  disease  is  really  anthrax  or 
not.  If  an  animal  has  died  suddenly  and  has  created  a  suspicion 
of  anthrax,  all  that  it  is  necessary  to  do  is  to  cut  cff  an  ear — or  a 
foot  in  the  case  of  a  sheep — and  make  a  cover-glass  preparation  at 
the  first  opportunity. 

A  farmer  with  a  case  of  anthrax  must  be  made  to  realise  the 
fact  that  an  enormous  quantity  of  poisonous  material  has  to  te 
dealt  with.  In  fact,  an  infected  animal  is  more  dangerous  when 
dead  than  alive.  The  owner  or  person  in  charge  must  immediately 
notify  to  a  police  constable  the  existence,  or  even  a  suspicion  of 
the  existence,  of  the  disease.  Prompt  measures  must  be  taken 
to  destroy  the  carcass  and  all  traces  of  the  blood,  and  thus  to 
reduce  to  a  minimum  the  chance  of  the  disease  spreading  to  the 
rest  of  the  stock,  and  of  creating  fresh  outbreaks  in  the  future. 
Every  possible  precaution  must  be  taken  to  prevent  the  blood  of 
the  dead  animal  from  contaminating  the  pasture,  byre,  or  water 
supply.  The  rest  of  the  stock  should  be  removed  from  the  pasture 
or  cowshed  where  the  disease  has  broken  out.  It  is  desirable  to 
give  a  complete  change  of  food  and  water,  and  the  whole  of  the 
stock  should  be  examined  every  day  for  a  week,  and  any  animals 
showing  a  rise  of  temperature  should  at  once  be  isolated  from  the 
rest.  Preventive  inoculation  has  been  recommended  to  protect 
the  rest  of  the  stock,  but  there  is  not  sufficient  evidence  of  the 
safety  of  the  process  to  lead  to  the  adoption  of  this  treatment. 
Animals  ready  for  the  butcher  may  be  removed  from  the  risk  of 
infection  by  immediate  slaughter.  To  disinfect  the  pasture  the 
best  plan  is  a  heavy  top-dressing  of  lime,  and  after  six  weeks 
stock  may  be  readmitted,  though  not  without  some  risk.  If 
year  after  year  cases  of  anthrax  occur  on  a  particular  pasture, 
the  most  obvious  precaution  is  to  keep  stock  from  it  altogether 
and  convert  it  into  arable  land.  As  roots  grown  on  anthrax- 
infected  soil  have  been  known  to  convey  the  disease,  the  wisest 
course  if  we  have  to  deal  with  a  small  field  or  comparatively 
small  tract  of  land  is  to  throw  it  out  of  cultivation  or  to  plant 
it  with  trees. 


ANTHRAX.  201 


DISPOSAL  OF  THE  CARCASS. 

The  surest  method  to  render  harmless  all  the  bacilli  which 
exist  in  the  carcass  is  burning,  but  cremation  offers  practical 
difficulties,  especially  if  several  carcasses  have  to  be  destroyed.  In 
the  case  of  an  animal  dying  in  a  town,  the  local  conditions  may 
render  it  best  to  adopt  destruction  by  burning  or  by  means 
of  chemicals.  In  such  a  case  the  carcass  should  be  covered 
with  quicklime,  and  then  taken,  in  charge  of  an  officer  of  the 
Local  Authority,  to  a  horse -slaughterer's  or  knacker's-yard,  and 
destroyed  by  exposure  to  a  high  temperature,  or  by  chemical  agents 
especially  in  the  vicinity  of  chemical  works.  Under  the  usual 
circumstances  of  death  occurring  on  a  farm,  fortunately  the  simple 
plan  of  burial,  with  the  addition  of  lime  or  other  chemical  agents, 
is  perfectly  efficacious,  and  even  without  the  use  of  chemicals  if  the 
carcass  has  been  left  unopened,  as  the  bacilli  die  rapidly  if  air  is 
excluded. 

Some  experiments  carried  out  by  M'Fadyean  clearly  indicate  the 
importance  of  leaving  the  carcass  unopened. 

On  July  16th  a  sheep  was  infected  with  anthrax  by  feeding  it  with  a 
virulent  culture.  Five  days  later  it  died,  and  a  microscopic  examina- 
tion of  blood  from  the  ear,  immediately  after  death,  showed  very  many 
anthrax  bacilli.  The  carcass  was  left  unskinned  and  unopened  until 
July  27th,  when  the  various  organs  were  cut  out  of  the  chest  and 
abdomen  and  placed  in  a  tin  box.  The  box  was  then  buried  at  a  depth 
of  about  two  feet  in  garden  earth,  and  left  there  undisturbed  until 
February  15th,  when  it  was  exhumed.  The  organs  had  become  con- 
verted into  adipocere,  and  this  was  thoroughly  mixed  up  with  water  and 
administered  to  a  sheep.  The  sheep  remained  perfectly  healthy.  In 
another  experiment  a  rabbit  was  inoculated  with  anthrax  on  June  1st. 
It  died  on  June  3rd,  and  blood  from  the  ear  contained  the  bacilli.  The 
rabbit  was  left  unopened  for  three  days,  and  then  placed  in  a  flower  pot 
and  buried  in  garden  earth  at  a  depth  of  two  feet;  It  was  exhumed  on 
February  15th.  The  tissues  were  all  destroyed  by  putrefaction,  and  the 
earth  in  contact  with  the  bones  was  administered  to  a  sheep  without 
conveying  the  disease  or  producing  any  ill  effects. 

Thus,  in  the  first  experiment,  the  lungs  and  the  intestines,  in 
which  spore  formation  was  most  likely  to  occur,  were  used  as  a 
test,  and  in  the  second  experiment  the  entire  carcass.  In  both  cases 
there  was  destruction  or  disappearance  of  the  bacilli,  and  these  tests, 
therefore,  confirm  in  a  very  marked  way  the  opinion  that  prompt 
burial  of  the  unopened  carcass  is  a  perfectly  safe  plan  to  adopt. 


202  INFECTIVE    DISEASES. 

If  an  animal  has  died  in  a  meadow,  a  pit  six  feet  deep  should 
be  dug  close  to  the  carcass,  and  if  quicklime  can  be  procured  with- 
out delay  the  carcass  should  be  buried  with  a  layer  about  a  foot  in 
depth  beneath  it  and  with  about  the  same  quantity  to  cover  it,  and 
the  pit  filled  up  with  the  excavated  soil. 

If  there  are  any  traces  of  blood  where  the  animal  lay,  the 
contaminated  ground  should  be  covered  with  quicklime  or  drenched 
with  strong  carbolic  acid,  and  the  whole  of  the  site  of  burial  fenced 
off  for  six  months.  If  an  animal  dies  near  a  brook  or  stream  then 
the  carcass  must  be  removed  for  burial  to  a  sufficient  distance  to 
prevent  any  reasonable  probability  of  contamination  of  the  water. 

If  death  has  occurred  in  the  byre,  the  carcass  must  be  removed 
to  the  nearest  and  most  convenient  spot  for  burial,  any  fodder  or 
litter  which  may  have  been  in  contact  with  the  deceased  animal  must 
be  destroyed,  and  the  shed  and  cart  and  any  utensils,  hurdles,  etc., 
disinfected.  For  the  latter  purpose  thorough  scouring  with  water 
and  then  washing  with  limewash  is  recommended.  The  limewash 
should  be  prepared  immediately  before  use,  and  four  ounces  of 
chloride  of  lime,  or  half  a  pint  of  commercial  carbolic  acid,  be  added 
to  each  gallon  of  limewash. 

The  following  is  an  illustration  of  the  value  of  preventive 
measures  based  upon  a  knowledge  of  the  exact  nature  of  the  disease. 
A  farm  on  the  banks  of  the  Yeo  was  repeatedly  attacked  by 
anthrax.  One  morning  two  sheep  died,  and  other  cases  followed. 
The  farmer  learnt  that  his  predecessor  had  buried  cattle  which  had 
died  of  anthrax  on  the  very  spot  where  the  sheep  were  folded.  He 
removed  his  flock,  and  had  no  further  losses  among  the  sheep,  but 
he  continued  to  lose  cattle  grazing  in  the  pastures  by  the  river. 
These  pastures  were  occasionally  flooded  by  the  Yeo.  Another 
farmer  in  the  same  locality  heavily  manured  a  field,  and  shortly 
afterwards  anthrax  broke  out  in  a  most  deadly  form  on  his  farm. 

What  was  the  cause  of  these  mysterious  outbreaks  ?  The 
explanation  was  forthcoming,  and  prevention  an  easy  matter.  The 
river  Yeo  received  the  washings  from  the  wool  factories  at  Yeovil, 
and  the  pastures  were  contaminated  by  anthrax  spores  in  the 
deposit  which  was  left  behind  when  the  flood  subsided.  In  the 
second  instance,  it  was  found  that  the  manure  used  for  dressing 
the  pasture  consisted  of  a  quantity  of  refuse  from  the  wool  factories. 

Infected  wool  from  foreign  countries  is  one  of  the  principal 
sources  of  the  disease  in  this  country,  and  the  remedy  is  to  insist 
upon  the  factories  destroying  their  refuse  instead  of  its  being 
allowed  to  contaminate,  the  rivers  or  to  be  sold  as  manure. 


ANTHRAX.  203 

So  long  as  this  source  of  the  disease  was  unknown  anthrax 
continued  to  be  spread  through  the  agency  of  the  wool  factories. 

Anthrax  spores  may  also  be  introduced  with  foreign  oats,  hay, 
and  manure,  so  that  it  is  almost  impossible  absolutely  to  prevent 
the  importation  of  the  disease;  but  the  danger  of  its  unlimited 
extension  and  disastrous  losses  can  be  minimised,  and  the  com- 
munication of  the  disease  to  man  and  to  swine  entirely  avoided  by 
simple  precautions. 

ANTHRAX  IN  SWINE. 

The  occurrence  of  anthrax  in  swine  is  a  subject  upon  which 
there  has  long  been  considerable  diversity  of  opinion.  Some  of  the 


FIG.  99. — ANTHRAX  IN  SWINE.  From  a  photograph  taken  during  life,  showing  a 
swollen  condition  of  the  neck  and  throat  six  days  after  ingestion  of  part  of  the 
viscera  of  a  bullock  which  had  died  from  anthrax. 

earliest  writers  on  the  diseases  of  animals  speak  of  outbreaks  of 
anthrax  among  swine,  but  whether  any  or  all  of  these  outbreaks 
were  examples  of  true  anthrax  has  long  been  a  matter  of  un- 
certainty; for  it  is  well  known  that  diseases  quite  distinct  were 
included  under  the  name  anthrax. 

Menschel  states  that  in  an  outbreak  in  which  twenty-four 
persons  were  attacked  with  malignant  pustule,  many  of  them  from 
eating  the  flesh  of  beasts  suffering  from  anthrax,  pigs  which  were 
fed  on  the  same  flesh  also  became  affected,  and  a  woman  who  ate 
some  of  the  diseased  pork  was  subsequently  ill. 

Roche-Lubin,  while  apparently  accepting  the  occurrence  of 
anthrax  in  swine,  taught  that  the  pig  resisted  inoculation  with  the 
blood  of  ^  different  species. 


204  INFECTIVE   DISEASES. 

In  this  country  accounts  have  been  published  from  time  to  time 
of  a  fatal  disease  in  pigs  induced  by  eating  the  flesh  of  animals 
which  had  died  of  what  was  described  as  "  blood-poisoning." 

Some  very  striking  cases  occurred  in  the  practice  of  Mr.  Wilson, 
of  Berkhampstead,  and  were  reported  in  the  Veterinarian.  A 
farmer  consulted  Mr.  Wilson  respecting  an  illness  with  which  his 
pigs  were  affected,  stating  that  two  or  three  were  dead  and  many 
others  seriously  ill.  They  were  strong  hogs,  ranging  from  six  to 
nine  months  old.  On  inquiry  it  was  ascertained  that  the  farmer 
had  lost  a  beast  suddenly  about  a  week  previously,  that  the  carcass 
had  been  opened  in  the  yard,  and  the  viscera  thrown  to  the  pigs. 
Mr.  Wilson  expressed  the  belief  that  the  disease  was  anthrax,  and 
stated  that  he  found  the  pigs  exhibiting  many  of  the  symptoms 
observable  in  cattle,  with  the  additional  one  of  enlargement  round 
the  throat  from  infiltration  of  a  yellow  fluid  causing  discoloration 
of  the  skin. 

Also,  in  the  reports  of  the  Agricultural  Department  of  the  Privy 
Council  thirteen  pigs  were  reported  as  suffering  from  anthrax  in 
1886,  and  one  hundred  and  fifty-nine  in  1887. 

But  the  question  arose  whether  the  disease  in  the  pigs  was 
genuine  anthrax  or  septic  poisoning. 

Williams  says :  "  The  flesh  of  animals  which  have  died  or  have 
been  killed  whilst  suffering  from  the  disease  [anthrax]  should  not 
be  used  as  food  either  for  men,  pigs,  or  dogs,  as  it  is  apt  to  cause 
death  by  blood  poisoning " ;  and  Steel  writes  :  "  Pigs,  dogs,  and 
poultry  should  not  be  allowed  to  feed  on  blood,  flesh,  and  eject  a 
of  anthrax  victims,"  but  no  statement  is  made  as  to  the  nature  of 
the  illness  produced.  No  doubt  these  writers  have  been  greatly 
influenced  by  the  opinion  of  many  bacteriologists,  for  Toussaint 
maintained  that  pigs  could  not  be  infected  with  anthrax,  and  a 
similar  view  was  at  one  time  upheld  in  this  country  by  Klein, 
who  stated  that  pigs  were  very  insusceptible.  In  Germany  also, 
pigs  have  been  credited  with  an  immunity  from  this  disease. 

In  the  face  of  these  conflicting  statements  the  author  carried 
out  a  series  of  experiments  in  order  to  ascertain  the  nature  of  the 
disease  in  swine  resulting  from  the  ingestion  of  the  offal  of  animals 
which  had  died  of  anthrax ;  and  the  result  of  inoculation  with  blood 
of  animals  which  had  died  of  anthrax,  and  with  pure  cultivations 
of  the  Bacillus  anthracis. 

As  a  result  of  these  experiments  genuine  anthrax  was  produced 
in  swine  (a)  by  feeding  them  with  anthrax  offal ;  (b)  by  injection 
of  blood  of  a  bullock  which  had  died  of  anthrax ;  (c)  by  passing 


ANTHRAX. 


205 


bacilli  through  the  guinea-pig,  and  transmitting  them  to  swine  by 
injection  of  blood  from  the  spleen ;  (d)  by  injecting  a  pure  cultiva- 
tion of  the  anthrax  bacillus;  (e)  and  lastly,  the  anthrax  bacillus 
was  isolated  from  swine  in  which  the  disease  was  accidentally  induced 
on  a  farm,  and  the  disease  reproduced  by  inoculation  of  guinea-pigs 
and  mice  with  blood  from  the  spleen. 

The  Author's  Conclusions. — Swine  of  all  ages  can  be  affected  with 
anthrax.  If  the  disease  is  induced  by  ingestion  of  anthrax  offal, 
the  tonsils  are  ulcerated,  and  constitute  the  point  of  access  of  the 
bacilli  to  the  blood.  In  such  cases  the  characteristic  symptom  is 


FIG.  100. — ANTHRAX  IN  SWINE.  From  a  photograph  taken  post-mortem.  Death 
occurred  four  days  after  the  ingestion  of  offal  from  a  bullock  which  had  died  of 
anthrax,  and  there  was  well-marked  oedema  of  the  throat,  cheeks,  and  eyelids. 

enormous  swelling  around  the  throat.  If  the  disease  is  induced  by 
hypodermic  injection,  the  same  cedematous  infiltration  of  the  tissues 
occurs  at  the  place  selected  for  inoculation.  Death  may  occur 
in  twenty-four  hours,  or  not  until  after  five  or  six  days.  There 
is  a  rapid  rise  of  temperature,  usually  a  rash -like  discoloration 
of  the  skin,  sometimes  loss  of  power  over  the  limbs,  and  general 
weakness  and  disinclination  to  move ;  the  a  nimal  may  lie  helplessly 
on  its  belly,  and  utter  plaintive  cries  when  disturbed.  At  the  post- 
mortem the  most  characteristic  feature  is  the  gelatinous  oedema 
which,  in  the  case  of  ingestion  of  offal,  is  found  around  the  throat. 
There  is  usually  congestion  of  all  the  organs  and  engorgement  of 


206  INFECTIVE    DISEASES. 

the  heart  and  large  vessels,  fluid  in  the  cavities  of  the  chest  and 
abdomen,  and  enlargement  and  haemorrhage  into  the  lymphatic 
glands.  There  is  in  some  cases  inflammation  of  the  intestines  with 
.submucous  and  subserous  haemorrhages.  The  spleen  may  be  normal 
in  size,  pale  and  flabby,  and  the  liver  only  slightly  congested  and 
friable ;  in  other  cases  the  condition  is  characteristic,  the  spleen 
is  the  seat  of  haemorrhage,  causing  more  or  less  local  enlargement, 
which  is  superficially  of  a  deep  purple  colour ;  the  liver  may  also 
be  greatly  congested,  very  friable,  and  marked  with  purple  patches. 
The  examination  of  the  blood  of  the  heart  and  spleen  for  anthrax 
bacilli  must  be  carried  out  with  great  perseverance  and  discrimi- 
nation, as  they  are  present  only  in  small  numbers,  and  in  some 
cases  have  given  place  entirely  to  septic  organisms.  Inoculation 
with  the  blood  will  produce  either  typical  anthrax,  or  malignant 
cedema  or  some  other  form  of  septicaemia.  Possibly  in,  the  cases 
arising  from  ingestion  of  offal  the  ulcerated  condition  of  the  throat 
affords  a  nidus  and  a  means  of  access  for  septic  organisms.  It 
is  also  well  known  that  blood  in  a  state  of  putrefaction  may 
contain  the  bacillus  of  malignant  cedema.  In  the  presence  of 
putrefactive  organisms  the  anthrax  bacillus  rapidly  disappears.  If, 
therefore,  inoculation  of  guinea-pigs  or  mice  is  used  as  a  test  for 
ascertaining  the  nature  of  an  outbreak  in  swine,  it  must  not  be 
concluded,  if  Pasteur's  or  some  other  form  of  septicaemia  result, 
that  the  disease  was  not  anthrax,  while,  on  the  other  hand,  the 
discovery  of  the  anthrax  bacillus  in  the  blood  of  the  pig,  or  the 
production  of  anthrax  in  guinea-pigs  or  mice,  is  positive  evidence 
as  to  the  nature  of  the  original  disease. 

Peuch,  in  France,  had  obtained  similar  results  by  injecting  pigs 
with  anthrax  blood  and  anthrax  cultures.  He  also  carried  out 
some  interesting  experiments  bearing  on  public  health.  The  leg  of 
a  pig  which  had  died  of  anthrax  was  covered  with  pounded  sea-salt. 
Previously  to  the  curing,  a  slice  of  the  flesh  was  squeezed  in  a  meat- 
press,  and  the  liquid  thus  obtained  was  employed  for  inoculation. 
The  animals  inoculated  died  of  typical  anthrax.  In  six  weeks  the 
curing  was  considered  to  be  completed,  and  a  slice  was  cut  from  the 
ham  and  soaked  in  filtered  water.  The  juice  was  extracted  in  the 
meat-press,  and  employed  for  the  inoculation  of  four  guinea-pigs 
and  three  rabbits.  Slight  swelling  and  a  certain  amount  of  redness 
at  the  seat  of  inoculation  were  the  only  results.  A  few  drops  of 
the  muscle-juice  were  added  to  sterilised  broth,  and  produced  a 
mixed  cultivation  of  micrococci  and  motile  bacilli.  A  rabbit  and  two 
guinea-pigs  inoculated  with  the  cultivation  remained  quite  healthy. 


ANTHRAX.  207 

These  experiments  demonstrated  that  salting  destroys  the  viru- 
lence of  the  flesh  of  pigs  which  have  died  of  anthrax,  but  in  order 
to  obtain  this  result  the  salting  must  be  thoroughly  carried  out.  If 
the  process  be  incomplete  the  flesh  is  still  virulent.  Thus  the  leg 
of  a  pig  salted  for  only  fourteen  days  furnished  a  juice  which 
possessed  a  certain  amount  of  virulence.  Out  of  three  inoculated 
rabbits,  one  died  in  ninety-seven  hours  of  anthrax,  and  the  others 
recovered.  Three  guinea-pigs  all  succumbed,  and  a  fourth  guinea- 
pig  inoculated  with  a  cultivation  from  the  muscle-juice  also  died. 
Peuch  considers  that  there  is  danger  in  consuming  flesh  which  has 
not  been  thoroughly  cured. 

As  it  has  been  clearly  shown  that  pigs  may  become  infected  with 
anthrax,  these  animals  come  under  the  Anthrax  Order  of  1886. 
This  provides  for  the  disposal  of  the  carcass;  and  although  Peuch 
has  shown  that  salting  destroys  the  virulence  of  the  flesh  of  pigs 
which  have  died  of  anthrax,  there  can  be  no  doubt  that  it  is  quite 
right  that  such  animals  should  be  condemned  as  unfit  for  food. 

Further,  the  recognition  of  the  occurrence  of  true  anthrax  in 
swine  is  an  additional  reason  for  condemning  the  Continental  practice 
of  eating  hams,  sausages,  etc.,  in  the  raw  state.  Indeed,  the  viru- 
lence of  anthrax  flesh  suggests  one  possible  explanation  of  some 
of  those  obscure  cases  of  meat  poisoning  which  have  occurred  in 
this  country.  It  is  possible  that  the  flesh  of  animals  which  had 
died  of  anthrax  was  used  in  the  preparation  of  sausages,  pork- pies, 
etc.,  and  that  the  cooking  was  not  sufficient  to  deprive  the  meat 
of  its  poisonous  properties. 

EQUINE  ANTHRAX. 

Veterinary  authorities  have  described  "  Anthrax  in  the  Horse," 
but  it  remains  to  be  seen  whether  there  are  not  two  or  more  affec- 
tions included  under  this  heading.  Fleming  says  :  "  The  most 
acute  form  of  anthrax,  the  apoplectic,  is  somewhat  rare  in  the 
horse,  and  has  perhaps  been  most  frequently  observed  on  the 
Continent.  Though  cases  are  recorded,  but  through  an  error  in 
diagnosis,  under  other  names  in  the  veterinary  literature  of  this 
country,  I  have  only  witnessed  two  cases  in  England ;  though  during 
the  intense  summer  heat  in  the  north  of  China  I  had  several." 

The  question  to  which  the  author  is  in  a  position  to  give  a 
definite  answer  is,  whether  the  disease  produced  by  the  Bacillus 
anthracis  ever  occurs  in  the  horse.  Whether  that  has  been  pre- 
viously determined,  at  any  rate  in  this  country,  it  is  difficult  to  say. 


208  INFECTIVE    DISEASES. 

Fleming  in  describing  the  pathological  anatomy  of  anthrax  in  the 
horse,  says  :  "  The  spleen  is  double  and  treble  its  ordinary  volume  ; 
its  surface  is  sometimes  bosselated  by  tumours;  its  texture  is 
softened  and  transformed  into  a  viscid  reddish-brown  or  violet  mass, 
and  the  mesenteric  glands  are  infiltrated.  The  blood  in  it  has 
been  found  to  contain  bacteridia  when  examined  soon  after  death." 
Williams,  who  says  that  "  anthrax  in  the  horse  rarely  occurs 
in  this  country,"  adds,  that  it  is  prevalent  in  India,  and  is 
there  termed  "  Loodiana  disease,"  and  in  Africa  "  Horse-sick- 
ness." But  "  Horse-sickness,"  from  recent  researches,  is  certainly 
not  anthrax.  Williams  described  a  case  which  occurred  in  1879  as 
one  of  anthrax.  A  carriage-horse  died  suddenly  while  in  harness  ; 
"  a  large  black  tumour  was  found  in  the  lungs,  and  the  pulmonary 
arteries  were  engorged  with  black  tarry  blood,  which,  when  micro- 
scopically examined,  was  found  to  contain  the  bacilli  in  a  most 
perfect  form,  and  very  numerous  indeed."  In  1884,  an  outbreak 
of  charbonous  fever  occurred  in  Liverpool.  Williams  proceeded 
to  investigate  the  outbreak,  and  found  two  horses  dead  on  his 
arrival,  one  having  died  only  a  few  hours  previously.  The  bacilli 
from  the  blood  in  this  case  are  figured,  and  the  following  statement 
made  :  "  These  bacilli  seem  to  differ  from  those  of  splenic  fever,  being 
rather  smaller  in  diameter,  and  so  far  as  my  observations  go, 
multiply  by  fission  only,  not  developing  spores." 

On  the  other  hand,  the  author  investigated  the  blood  of  a  mare 
which  was  supposed  to  have  died  of  anthrax,  and  on  examining 
cover-glass  preparations  of  the  blood,  it  was  found  to  contain  large 
numbers  of  bacilli  with  the  characteristic  microscopical  appearances 
of  anthrax  bacilli.  To  place  the  question  beyond  any  possible  doubt 
a  number  of  tubes  of  agar-agar  were  inoculated.  These,  after  three 
days  in  the  incubator,  produced  typical  cultivations,  and  on  examina- 
tion by  the  ordinary  methods  and  by  double-staining,  yielded  very 
beautiful  preparations  of  filaments  and  spores. 

At  the  same  time  that  the  cultivations  were  prepared,  two  mice 
were  inoculated  at  the  root  of  the  tail  with  a  trace  of  the  blood. 
Two  days  afterwards  they  were  both  found  dead,  and  with  the 
characteristic  post-mortem  appearances,  spleen  much  enlarged,  and 
anthrax  bacilli  in  enormous  numbers. 

There  can  be  no  doubt  that  true  anthrax  occurs  in  the  horse  ; 
and  the  author,  in  1887,  recommended  that  it  should  be  scheduled 
under  the  Contagious  Diseases  (Animals)  Act,  and  equine  anthrax 
has  been  included  in  the  Anthrax  Order  of  1895. 

More  recently  Pemberthy  has  described  cases  of  equine  anthrax 


ANTHRAX. 


209 


which  he  believes  to  have  been  the  result  of  infection  from  feeding 
on  foreign  oats  or  imported  hay. 

Preventive  Inoculation. — The  prevention  of  anthrax  by 
means  of  protective  inoculation  or  vaccination  has  been  attempted 
on  a  very  large  scale  in  France,  and  it  is  claimed  that  the  results 
have  been  very  beneficial  to  agriculture  in  that  country  : — 


Animals 

Mortality. 

Total 

Na.  of 

Vacci- 

Total 

Average 

No.  of 

Veteri- 

nated 

loss 

Ios7 

Year. 

Animals 
Vacci- 
nated. 

nary 
Re- 
ports. 

after 
Receipt 
of 
Reports. 

After 
1st 
Vacci- 
nation. 

After 
2nd 
Vacci- 
nation. 

During 
rest  of 
Year. 

Total. 

per 
cent. 

before 
Vacci- 
nation. 

/1S82 

270..040 

112 

243,199 

756 

847 

1,037 

2,640 

1-08 

10% 

1883 

268,505 

103 

193,119 

436 

272 

784 

1,492 

0-77 

1884 

316,553 

109 

231,693 

770 

444 

1,083 

2,247 

0-97 

1885 

342,040 

144 

280,107 

884 

735 

990 

2,609 

0-93 

1886 

313,288 

88 

202,064 

652 

303 

514 

1,469 

0-72 

1887 

293,572 

107 

187,811 

718 

737 

968 

2,423 

1-29 

Sheep  ^ 

1888 

269,574 

50 

101,834 

149 

181 

300 

630 

0-62 

1889 

239,974 

43 

88,483 

238 

285 

501 

1,024 

1-16 

1890 

223,611 

69 

69,865 

331 

261 

244 

836 

1-20 

1891 

218,629 

65 

53,640 

181 

102 

77 

360 

0-67 

1892 

259,696 

70 

63,125 

319 

183 

126 

628 

0-99 

,1893 

281,333 

30 

73,939 

234 

56 

224 

514 

0-69 

Total     . 

3,296,815 

990 

1,788,677 

5,668 

4,406 

6,798 

16,872 

0-94 

(0-32%) 

(0-24%) 

(0-38%) 

/1882 

35,654 

127 

22,916 

22 

12 

48 

82 

0-35 

5% 

1883 

26,453 

130 

20,501 

17 

1 

46 

64 

0-31 

1884 

33,900 

139 

22,616 

20 

13 

52 

85 

0-37 

1885 

34,000 

192 

21,073 

32 

8 

67 

107 

0-50 

1886 

39,154 

135 

22,113 

18 

7 

39 

64 

0-29 

Oxen 

1887 

48,484 

148 

28,083 

23 

18 

68 

109 

0-39 

or 

1888 

34,464 

61 

10,920 

8 

4 

35 

47 

0-43 

Cows 

1889 

32,251 

68 

11,610 

14 

7 

31 

52 

0-45 

1890 

33,965 

71 

11,057 

5 

4 

14 

23 

0-21 

1891 

40,736 

68 

10,476 

6 

4 

4 

14 

0-13 

1892 

41,609 

71 

9,757 

8 

3 

15 

26 

0-26 

,1893 

38,154 

45 

9,840 

4 

1 

13 

18 

0-18 

438,824 

1,255 

200,962 

177 

82 

432 

691 

0-34 

(0-09%) 

(0-04%) 

(0-21%) 

i 

14 


210 


INFECTIVE    DISEASES. 


The  vaccine  is  supplied,  by  a  company  in  Paris,  in  two  strengths. 
Reports  are  supplied  by  veterinary  surgeons,  and  the  results  have 
been  tabulated  by  Chamberland  and  published,  and  commented  upon 
by  Cope  in  a  report  to  the  Board  of  Agriculture  (1894).  The  column 
of  deaths,  in  the  above  table,  includes  the  animals  which  died  from 
the  vaccination,  and  those  which  died  from  natural  infection. 

It  is  claimed  that  the  percentage  of  losses  has  been  reduced  from 
10  per  cent,  to  "94  per  cent,  in  sheep,  and  from  5  per  cent,  to  '34  per 
cent,  in  cattle.  Cope,  in  the  report  just  referred  to,  regards  these 
conclusions  as  somewhat  fallacious,  because  in  order  to  prove  that 
the  animals  inoculated  received  immunity,  it  should  be  shown  that 
they  were  subsequently  exposed  to  the  risks  of  natural  infection. 
This  was  not  the  case.  But  a  report  obtained  from  the  Bureau  in 
Paris  gives  the  actual  number  of  animals  on  each  of  the  infected 
farms,  and  the  number  which  have  died  of  the  disease  ;  and  when 
compared  with  Chamberland's  statistics  it  is  evident  that  nine-tenths 
were  not  on  farms  where  the  disease  appeared — at  least,  during 
1889-92 — and  that  the  deaths  from  anthrax  on  those  farms  where 
it  was  reported  to  exist  were,  if  anything,  higher  than  they  were 
supposed  to  be  prior  to  the  introduction  of  the  system  of  vaccination  ; 
and  in  spite  of  the  immense  number  of  animals  vaccinated  the 
official  returns  obtained  from  Paris,  by  Cope,  indicate  that  the 
mortality  from  anthrax,  calculated  in  the  ordinary  way,  remains  as 
high  as  ever. 

Anthrax  in  France. 


Year. 

No.  of 
Outbreaks  re- 
ported. 

No.  of  Animals 
in  Premises. 

No.  of  which 
Died. 

Percentage  of 
Loss. 

1889 

618 

22,599 

1,458 

6-5 

1890 

536 

'  24,073 

1,123 

4-7 

1891 

570 

21,356 

1,444 

6-8 

1892 

607 

28,199 

1,581 

5-6 

CATTLE. 

1889 

— 

6,059 

700 

11-6 

1890 

— 

5,365 

771 

14-4 

1891 

— 

7,299 

849 

11-6 

1892 

— 

5,058 

804               15-9 

SHEEP. 

1889 

16,540 

755                 4-6 

1890 

18,708' 

352                  1-Sl 

1891                                         14,057 

545 

4-2 

1892 

—                   23,141 

777 

3-4 

ANTHRAX.  211 

In  Germany,  veterinary  and  agricultural  authorities  agree  that 
the  results  have  not  met  with  the  success  which  has  been  claimed 
for  vaccination  in  France.  Experiments  were  undertaken  for  the 
German  Government,  and  in  one  set  of  experiments  twenty-five  sheep 
were  vaccinated  with  the  first  vaccine  without  an  accident,  but  three 
died  five  days  after  the  second  vaccine.  In  another  experiment  two 
hundred  and  fifty-one  sheep  were  vaccinated  with  only  one  death, 
and  subsequent  inoculation  with  virulent  anthrax  proved  that  they 
had  immunity. 

Six  head  of  cattle  were  vaccinated  without  any  loss,  and  six  more 
were  used  for  a  control  experiment.  Inoculation  with  virulent  virus 
proved  fatal  to  the  control  animals,  but  the  vaccinated  were  pro- 
tected. These,  with  other  animals  similarly  vaccinated,  amounting 
in  all  to  two  hundred  and  sixty-six  sheep  and  eighty-three  head  of 
cattle,  were  then  turned  out  to  graze  on  infected  pastures  with  two 
hundred  and  sixteen  unvaccinated  sheep  as  a  control  experiment. 
Within  five  months  four  of  the  vaccinated  and  eight  of  the  un- 
vaccinated sheep  died  of  anthrax,  and  one  of  the  vaccinated  and  one 
of  the  unvaccinated  cattle. 

The  result  of  these  experiments  led  to  the  following  conclusions  : — 

(1)  That  the  first  vaccine  is  mild  and  harmless. 

(2)  That   the  second  vaccine,  even  in  the  hands  of  experts,  is 
dangerous  and  often  fatal. 

(3)  That  sheep  are  more  affected  than  cattle  by  the  injections,, 
exhibiting  fever  and  other  indications  of  illness. 

(4)  That  cattle  and  sheep  which  recover  from  the  vaccination 
have  an  immunity  against  anthrax  when  tested  by  experimental 
inoculation. 

(5)  That   vaccinated   cattle   and   sheep   tested    by   exposure   to 
natural   infection    by   grazing   on   infected   pastures    contract   the 
disease  in  the  ordinary  way. 

(6)  That  the  time  for  which  immunity  is  conferred  has  not  been 
determined. 

In  England,  Klein  tested  the  vaccine,  with  the  residt  that  animals 
either  succumbed  to  the  vaccine,  or  to  virulent  anthrax  after  recovery 
from  the  vaccine.  Protective  inoculation  has  also  been  employed  in 
a  few  instances  by  leading  agriculturists,  but  with  very  unsatis- 
factory results. 

Stamping -out  System. — In  Germany  the  conclusion  is  that 
the  safest  measures  are  destruction  of  carcasses  and  disinfection,  and 
that  inoculation  will  have  no  effect  in  lessening  the  loss  caused  by 
this  disease. 


212  INFECTIVE    DISEASES. 

Iii  England  the  stamping- out  system  has  been  advocated  for 
many  years,  and  is  still  regarded  as  the  only  reliable  means  for 
suppressing  the  disease ;  and  the  possible  introduction  of  the  disease 
among  healthy  stock  by  vaccination,  and  especially  in  localities  in 
which  anthrax  is  unknown,  would  be  contrary  to  the  principles  upon 
which  the  system  is  based.  These  principles  are  illustrated  by  the 
following  extracts  from  the  Anthrax  Order  of  1895  : — 

NOTIFICATION. 

2. — (1)  Every  person  having  or  having  had  in  his  possession  or  under 
his  charge,  an  animal  affected  with  or  suspected  of  anthrax,  shall,  with  all 
practicable  speed,  give  notice  of  the  fact  of  the  animal  being  so  affected  or 
suspected,  to  a  constable  of  the  police  force  for  the  police  area  wherein 
the  animal  so  affected  or  suspected  is  or  was. 

(2)  The  constable  shall  forthwith  give  information  of  the  receipt  by 
him  of  the  notice  to  an  Inspector  of  the  Local  Authority,  who  shall  forth- 
with report  the  same  to  the  Local  Authority. 

(3)  The   Inspector    of    the   Local    Authority  shall    forthwith    give 
information  of  the  receipt  by  him  of  the  notice  to  the  Medical  Officer 
of  Health  of  the  Sanitary  District  in  which  the  affected  or  suspected 
animal  is  or  was. 

Duty  of  Inspector  to  act  immediately. 

3.  An  Inspector  of  a  Local  Authority  on  receiving  in  any  manner 
whatsoever  information  of  the  supposed  existence  of  anthrax,  or  having 
reasonable  ground  to  suspect  the  existence  of  anthrax,  shall  proceed  with 
all  practicable  speed  to  the  place  where  such  disease,  according  to  the 
information  received  by  him,  exists,  or  is  suspected  to  exist,  and  shall 
there  and  elsewhere  put  in  force  and  discharge  the  powers  and  duties 
conferred  and  imposed  on  him  as  Inspector,  by  or  under  the  Act  of  1894 
and  this  Order. 

Public  Wanting  as  to  Existence  of  Disease. 

4. — (1)  The  Local  Authority  may,  if  they  think  fit,  give  public 
warning  by  placards,  advertisement,  or  otherwise,  of  the  existence  of 
anthrax  in  any  shed,  stable,  building,  field,  or  other  place,  with  or  without 
any  particular  description  thereof,  as  they  think  fit,  and  may  continue  to 
do  so  during  the  existence  of  the  disease,  and,  in  case  of  a  shed,  stable, 
building,  or  other  like  place,  until  the  same  has  been  cleansed  and  dis- 
infected in  accordance  with  this  Order. 

(2)  It  shall  not  be  lawful  for  any  person  (without  authority  or 
excuse)  to  remove  or  deface  any  such  placard. 

Milk  of  Diseased  or  Suspected  Cow  not  to  be  Removed. 

5.  Where  anthrax  exists  or  has  existed  in  any  shed,  stable,  building, 
or  other  place,  it  shall  not  be  lawful  to  remove  from  such  shed,  stable, 


ANTHRAX.  213 

building,  or  other  place  the  milk  of  any  cow  which  is  affected  with  or 
suspected  of  anthrax. 

Removal  of  Dung  or  other  Things. 

6.  It  shall  not  bei  lawful  for  any  person  to  send  or  carry,  or  cause  to 
be  sent  or  carried,  on  a  railway,  canal,  river,  or  inland  navigation,  or  in 
a  coasting  vessel,  or  on  a  highway  or  thoroughfare,  any  dung,  fodder,  or 
litter  that  has  been  in  any  place  in  contact  with  or  used  about  a  diseased 
or  suspected  animal,  except  with  a  Licence  of  the  Local  Authority  for 
the  District  in  which  such  place  is  situate,  on  a  certificate  of  an  Inspector 
of  the  Local  Authority  certifying  that  the  thing  moved  has  been,  so  far 
as  practicable,  disinfected. 

Disposal  of  Carcasses. 

1. — (1)  The  carcass  of  an  animal  which  at  the  time  of  its  death  was 
affected  with  or  suspected  of  anthrax  shall  be  disposed  of  by  the  Local 
Authority  as  follows  : — 

(i.)  Either  the  Local  Authority  shall  cause  the, carcass  to  be  buried 
as  soon  as  possible  in  its  skin  in  some  convenient  or  suitable  place 
removed  from  any  dwelling  house  and  at  such  a  distance  from  any 
well  or  watercourse  as  will  preclude  any  risk  of  the  contamination 
of  the  water  therein,  and  at  a  depth  of  not  less  than  six  feet 
below  the  surface  of  the  earth,  having  a  layer  of  lime  not  less 
than  one  foot  deep  beneath,  and  a  similar  layer  of  lime  above, 
the  carcass  ; 

(ii.)  Or  the  Local  Authority  may,  if  authorised  by  Licence  of  the 
Board,  cause  the  carcass  to  be  destroyed,  under  the  inspection  of 
the  Local  Authority,  in  the  mode  following:  The  carcass  shall 
be  disinfected,  and  shall  then  be  taken,  in  charge  of  an  officer  of 
the  Local  Authority,  to  a  horse-slaughterer's  or  knacker's-yard 
approved  for  the  purpose  by  the  Board,  or  other  place  so  approved, 
and  shall  be  there  destroyed  by  exposure  to  a  high  temperature, 
or  by  chemical  agents. 

(2)  With  the  view  to  the  execution  of  the  foregoing  provisions  of  this 
Article  the  Local  Authority  may  make  such  Regulations  as  they  think  fit 
for  prohibiting  or  regulating  the  removal  of  carcasses,  or  for  securing  the 
burial  or  destruction  of  the  same. 

(3)  Before  a  carcass  is  removed  for  burial  or  destruction  under  this 
Article  it  shall  be  covered  with  quicklime.     In  no  case  shall  the  skin  of 
the  carcass  be  cut,  nor  shall  anything  be  done  to  cause  the  effusion  of 
blood. 

(4)  A  Local  Authority  may  cause  or  allow  a  carcass  to  be  taken  into 
the  District  of  another  Local  Authority  to  be  buried  or  destroyed,  with 
the  previous  consent  of  that  Local  Authority,  but  not  otherwise. 

Digging  Up. 

8.  It  shall  not  be  lawful  for  any  person,  except  with  the  Licence  of 
the  Board  or  permission  in  writing  of  an  Inspector  of  the  Board,  to  dig 


214  INFECTIVE    DISEASES. 

up.  or  cause  to  be  dug  up,  the   carcass  of  any  animal    that   has   been 
buried. 

Disinfection  in  Case  of  Anthrax. 

9. — (1)  The  Local  Authority  shall  at  their  own  expense  cause  to  be 
cleansed  and  disinfected  in  the  mode  provided  by  this  Article — 

(a)  All  those  parts  of  any  shed,  stable,  building,  or  other  place   in 
which  a  diseased  or  suspected,  animal  has  been  kept  or  has  died  or 
been  slaughtered  ; 

(b)  Every  utensil,  pen,  hurdle,  or  other  thing  used  for  or  about  any 
diseased  or  suspected  animal ; 

(c)  Every  van,  cart,  or  other  vehicle  used  for  carrying  any  diseased 
or  suspected  animal  on  land  otherwise  than  on  a  railway. 

(2)  The  mode  of  the  cleansing  and  disinfection  of  such  shed,  stable, 
building,  or  other  place,  or  the  part  thereof,  shall  be  as  follows  : — 

(i.)  All  those  parts  aforesaid  of  the  shed,  stable,  building,  or  other 
place  shall  be  swept  out,  and  all  litter,  dung,  or  other  thing  that 
has  been  in  contact  with,  or  used  about,  any  diseased  or  suspected 
animal  shall  be  effectually  removed  therefrom  ;  then 

(ii.)  The  floor  arid  all  other  parts  of  the  shed,  stable,  building,  or  other 
place  with  which  the  diseased  or  suspected  animal  or  its  droppings 
or  any  discharge  from  the  mouth  or  nostrils  of  the  animal  has  come 
in  contact,  shall  be,  so  far  as  practicable,  thoroughly  washed  or 
scrubbed  or  scoured  with  water  ;  then 

(iii.)  The  same  parts  of  the  shed,  stable,  building,  or  other  place  shall 
be  washed  over  with  limewash  made  of  freshly  burnt  lime  and 
water,  and  containing  in  each  gallon  of  limewash  four  ounces  of 
chloride  of  lime  or  half  a  pint  of  commercial  carbolic  acid,  the 
limewash  being  prepared  immediately  before  use  ; 

(iv.)  Except  that  where  any  place  as  aforesaid  is  not  capable  of  being 
so  cleansed  and  disinfected,  it  shall  be  sufficient  if  such  place  be 
cleansed  and  disinfected  so  far  as  practicable. 

(3)  The  mode  of  the  cleansing  and  disinfection  of  such  utensil,  pen, 
hurdle,  or  other  thing,  and  such  van,  cart,  or  other  vehicle  aforesaid,  shall 
be  as  follows  : — 

(i.)  Each  utensil,  pen,  hurdle,  or  other  thing,  van,  cart,  or  other 
vehicle,  shall  be  thoroughly  scraped,  and  all  litter,  dung,  sawdust, 
or  other  thing  shall  be  effectually  removed  therefrom  ;  then 

(ii.)  It  shall  be  thoroughly  washed  or  scrubbed  or  scoured  with  water  ; 
then 

(iii.)  It  shall  be  washed  over  with  limewash  made  of  freshly  burnt 
lime  and  water,  and  containing  in  each  gallon  of  limewash  four 
ounces  of  chloride  of  lime  or  half  a  pint  of  commercial  carbolic 
acid,  the  limewash  being  prepared  immediately  before  use. 

(4)  All  litter,  dung,  or  other  thing  that  has  been  removed  from  any 
such  shed,  stable,  building,  place,  van,  cart,  or  vehicle  as  aforesaid,  shall 
be  forthwith  burnt  or  otherwise  destroyed  or  disinfected  to  the  satisfac- 
tion of  an  Inspector  of  the  Local  Authority. 


ANTHRAX.  215 

(5)  The  Local  Authority  may  make  such  Regulations  as  they  think 
fit  for  the  purpose  of  carrying  out  the  provisions  of  this  Article. 

Occupiers  to  give  Facilities  for  Cleansing. 

10. — (1)  Where  the  power  of  causing  any  place,  thing,  or  vehicle  to 
be  cleansed  and  disinfected  under  this  Order  is  exercised  by  a  Local 
Authority,  the  owner  and  occupier  and  person  in  charge  of  the  place, 
thing,  or  vehicle  shall  give  all  reasonable  facilities  for  that  purpose. 

(2)  Any  person  failing  to  comply  with  the  provisions  of  this  Article 
shall  be  deemed  guilty  of  an  offence  against  the  Act  of  1894. 

Regulations  of  Local  Authority  as  to  Movement  of  Animals, 
Fodder,  etc. 

11.  A  Local  Authority  may  make  such  Regulations  as  they  think  fit 
for  the  following  purposes,  or  any  of  them  : — 

(a)  For  prohibiting  or  regulating  the  movement  of  any  diseased  or 

suspected  animal  into  or  out  of  any  shed,  stable,  building,  field,  or 

other  place,  or  any  part  thereof  ; 

(6)  For  prohibiting  or  regulating  the  movement  of  any  animal  into 
or  out  of  any  shed,  stable,  building,  field,  or  other  place,  or  any 
part  thereof,   in   which  there   is  or    has   been   any  diseased    or 
suspected  animal  ;  and 

(c)  For  regulating  the  removal  out  of  any  shed,  stable,  building, 
field,  or  other  place  of  any  fodder,  litter,  or  other  thing  that  has 
been  in  contact  with  or  used  for  or  about  any  diseased  or  suspected 
animal  ; 

but  nothing  in  any  such  Regulation  shall  authorise  movement  in 
contravention  of  any  provision  of  any  Order  of  the  Board  for  the  time 
being  in  force  ;  and  a  Regulation  under  paragraph  (6)  of  this  Article 
shall  operate  so  long  only  as  any  animal  which  in  the  judgment  of  the 
Local  Authority  is  diseased  or  suspected  remains  in  the  shed,  stable, 
building,  field,  or  other  place  to  which  the  Regulation  refers,  and,  in  case 
of  a  shed,  stable,  building,  or  other  like  place,  until  the  same  has  been 
cleansed  and  disinfected  in  accordance  with  this  Order. 

Slaughter  hi  Anthrax  and  Compensation. 

12. — (1)  A  Local  Authority  may  if  they  think  fit  cause  to  be 
slaughtered — 

(a)  Any  animal  affected  with  anthrax  or  suspected  of  being  so 
affected  ;  and 

(6)  Any  animal  being  or  having  been  in  the  same  field,  shed,  or  other 
place,  or  in  the  same  herd  or  flock  or  otherwise  in  contact  with 
animals  affected  with  anthrax,  or  being  or  having  been  in  the 
opinion  of  the  Local  Authority  in  any  way  exposed  to  the  infection 
of  anthrax. 

(2)  The  slaughter  of  animals  under  this  Article  shall  be  conducted 
in  such  mode  as  will  so  far  as  possible  prevent  effusion  of  blood. 


216  INFECTIVE   DISEASES. 

(3)  The  Local  Authority  shall  out  of  the  local  rate  pay  compensation 
as  follows  for  animals  slaughtered  under  this  Article  : — 

(a)  Where  the  animal  slaughtered  was  affected  with  anthrax  the 
compensation  shall  be  one-half  of  the  value  of  the  animal 
immediately  before  it  became  so  affected  ;  and 

(5)  In  every  other  case  the  compensation  shall  be  the  value  of  the 
animal  immediately  before  it  was  slaughtered. 

(4)  Provided,  that  if  the  owner  of  the  animal  gives  notice  in  writing 
to  the  Local  Authority,  or  their  Inspector  or  other  officer,  that  he  objects 
to  the  animal  being  slaughtered,  it  shall  not  be  lawful  for  the  Local 
Authority  to  cause  that  animal  to  be  slaughtered  except  with  the  further 
special  authority  of  the  Board  first  obtained. 

Keeping  of  Swine  in  Slaughter-houses . 

16.  It  shall  not  be  lawful  for  any  person,  in  any  case  in  which  the 
slaughter  of  any  animal  is  authorised  or  required  by  this  Order,  to  use 
for  such  slaughter  any  slaughter-house  in  which  swine  are  kept. 

Whether  an  anthrax  virus  can  be  obtained  which  is  absolutely 
incapable  of  creating  centres  of  infection,  and  can  therefore  be 
recommended  with  safety  for  vaccination  as  an  auxiliary  and 
voluntary  measure,  is  a  matter  for  further  investigation. 


CHAPTER   XV. 

QUARTER-EVIL. — MALIGNANT  (EDEMA. — RAG-PICKERS'  SEPTICAEMIA. — 
SEPTICLEMIA   OF   GUINEA-PIGS.— SEPTICAEMIA   OF  MICE. 

QUARTER-EVIL  in  cattle,  malignant  oedema,  and  rag-pickers'  septicaemia 
in  man,  septicaemia  in  guinea-pigs,  and  septicaemia  in  mice,  are  all 
varieties  of  septicaemia  produced  by  bacilli. 

An  account  of  quarter-evil,  malignant  oedema,  and  rag-pickers' 
septicaemia  may  appropriately  follow  the  chapter  on  anthrax,  as 
they  have  certain  similarities  to  that  disease.  They  are,  however, 
not  only  distinct  from  anthrax,  but  must  be  carefully  distinguished 
from  each  other.  In  connection  with  these  forms  of  bacillary 
septicaemia  in  man  and  cattle  we  may  study  bacillary  septicaemia 
in  small  animals. 

QUARTER- EVIL. 

The  disease  known  in  this  country  as  quarter-evil  or  black-leg 
is  identical  with  the  French  Charbon  symptomatique  and  the 
German  Rauschbrand.  Symptomatic  anthrax  in  a  very  slight  degree 
resembles  anthrax.  The  disease  occurs  usually  in  young  cattle  from 
a  few  weeks  to  about  twelve  months  old,  and  attacks  sheep  and 
horses,  but  not  swine  or  poultry.  It  is  characterised  by  the  develop- 
ment of  an  emphysematous  swelling  of  the  subcutaneous  tissue  and 
muscles,  generally  over  the  hind  quarter.  Infected  animals  cease 
feeding,  the  temperature  rises,  lameness  supervenes,  and  death 
occurs  in  about  forty-eight  hours.  The  tumour  on  incision  is  found 
to  contain  a  quantity  of  dark  sanguineous  fluid,  with  characteristic 
bacilli. 

Bacillus  of  Quarter-evil  (Bacille  du  charbwi  symptomatique, 
Rauschbrand  bacillus).- — Motile  rods  with  rounded  ends,  3  to  5  p.  in 
length,  -5  to  *6  p.  in.  breadth.  Spore-formation  present.  The 
spores  are  oval,  generally  situated  near  the  extremity  of  the  rods, 
and  when  fully  developed  considerably  exceed  the  rods  in  diameter. 

217 


218 


INFECTIVE    DISEASES. 


Involution  forms  are  freely  developed  in   old  cultures,   and  in 
cultures  made  in  unsuitable  media.      The  bacilli  possess  numerous 

flagella,       and       their 

power  of  movement  at 

<0fft  once  distinguishes  them 

from    anthrax    bacilli. 

0  4?  They  can  be  cultivated 

1  «  in  the  ordinary  media 

o  OM»       P  in  the  absence  of  oxy- 

^  gen,  but  more  readily 

with    the    addition    of 
grape-sugar  or  glyce- 
rine.     Radiating  fila- 
f  9"        ments  grow   out  from 
^         the  more  or  less  spher- 
ical    colonies    directly 


/    o       -* 


V 

•>'* 


FIG.  101.     BACILLI  OF  QUAKTER-EVIL  x  1000.     From 
an  agar  culture  (FRANKEL  and  PFEIFFER). 


liquefaction  commences.  In  the  depth  of 
nutrient  gelatine  the  growth  occurs  in  two 
or  three  days  at  20°  to  25°  0.  towards  the 
lower  part  of  the  track  of  the  inoculating 
needle.  The  gelatine  slowly  liquefies,  and 
there  is  considerable  formation  of  gas  with 
the  development  of  a  peculiar  odour.  Spore- 
formation  occurs  freely  in  cultures,  but  not 
in  the  blood  of  infected  animals  until  after 
death. 

Guinea-pigs  inoculated  with  a  pure- 
culture,  or  with  spore-bearing  threads,  die 
in  twenty-four  to  thirty-six  hours.  An  em- 
physematous  infiltration  with  sanguineous 
serum  is  produced  at  the  seat  of  inoculation, 
and  the  surrounding  muscles  are  of  a  dark 
colour.  The  internal  organs  are  more  or  less  Fl(,  m  PURE_CULTURE  OF 
congested.  The  bacilli  are  found  in  the  BACILLI  OF  QUARTER-EVIL 

local    exudation    and    in    the    surrounding       IN   GRAPE-SUGAR  GELA- 
TINE      (FRANKEL 
PFEIFFER). 


tissue,    and    some     hours 


surrounding 
after    death    in 


and 


QUARTER-EVIL.  219 

increasing  numbers  in  the  blood  of  the  heart  and  in  the  internal 
organs. 

Quarter-evil  and  malignant  oedema,  though  possessing  points  of 
resemblance,  are  distinct  diseases.  Not  only  do  the  bacilli  in  the 
two  cases  differ  in  minute  morphological  and  biological  details,  but 
Kitasato  showed  that  guinea-pigs  rendered  immune  against  virulent 
quarter-evil  had  no  immunity  against  malignant  oedema. 

Protective  Inoculation. — Arloing,  Cornevin  and  Thomas  have 
produced  immunity  by  inoculating  healthy  cattle  with  a  small 
quantity  of  the  fluid  from  the  tumour  of  an  infected  animal. 
Recovery  takes  place,  and  subsequent  inoculation  with  a  strong  dose 
is  without  effect.  Similar  results  may  be  obtained  by  intravenous 
injection  of  a  few  drops  of  the  exudation.  For  general  application 
of  the  system  of  protective  inoculation,  the  virulent  liquid  and 
affected  muscles  are  dried  at  32°  to  35°  C.,  and  the  dried  mass 
triturated  with  water  and  heated  to  100°C.  This  is  used  as  the 
first  vaccine. 

An  infusion  similarly  prepared,  but  only  heated  to  80°  C.,  forms 
the  second  vaccine.  The  dry  powder  is  a  convenient  form  for 
general  distribution,  and  -^  of  a  gramme  is  triturated  with  5  cc. 
of  water,  and  |  cc.  is  injected  into  each  animal.  In  about  ten  days 
the  second  vaccine  is  employed,  and  cattle  so  treated  are  said  to 
have  a  complete  immunity  from  fatal  doses. 

The  place  chosen  for  the  injection  is  the  under  surface  of  the 
tail,  a  short  distance  from  the  extremity.  The  hair  is  clipped  at 
this  spot,  and  the  point  of  a  syringe  is  pushed  in  between  the  skin 
and  the  bone,  and  the  vaccine  slowly  injected. 

Eoux  and  Chamberland  produced  immunity  by  inoculation  of 
filtered  cultures.  Cultures  in  broth  were  deprived  of  bacilli  by 
heating  to  115°  C.,  or  by  filtration  through  porcelain.  Guinea-pigs 
were  inoculated  with  three  doses  of  30  cc.  at  intervals  of  two  days, 
and  subsequently  injected  with  a  solution  of  virulent  black-leg 
powder  and  lactic  acid,  which  killed  control  animals  in  twenty-four 
hours.  Kitt  employed  cultures  on  agar  a  fortnight  old,  or  fresh 
cultures  sterilised  by  steam  for  thirty  minutes.  It  was  found 
possible  to  confer  immunity  in  oxen,  sheep,  and  guinea-pigs  against 
the  most  virulent  extract.  Kitt's  method  has  the  advantage  over 
others  of  only  necessitating  one  single  injection.  Whether  these 
experiments  are  of  scientific  interest  rather  than  of  practical  value 
may  be  regarded  as  an  open  question. 

On  the  Continent,  and  especially  in  France,  vaccination  against 
quarter-evil  has  been  carried  out  extensively ;  and  by  comparing  the 


220  INFECTIVE    DISEASES. 

mortality  among  the  vaccinated  and  unvaccinated  in  localities  where 
the  disease  commonly  occurs,  it  has  been  said  that  the  results  are 
extremely  favourable.  The  matter  was  investigated  in  this  country 
by  a  committee  of  the  Midland  Veterinary  Medical  Association, 
and  in  the  course  of  the  experiments  some  surprising  results  were 
obtained.  Six  calves  and  four  sheep  were  vaccinated,  and  five 
calves  and  two  sheep  were  left  unvaccinated  as  a  control  experiment. 
The  seventeen  animals  were  subsequently  inoculated  with  virulent 
virus  in  the  form  of  dried  and  powdered  muscle.  In  forty- eight 
hours  all  the  sheep  died,  and  all  the  calves  exhibited  a  swelling  at 
the  seat  of  inoculation.  In  another  set  of  experiments,  healthy 
calves  inoculated  with  fresh  juice  from  the  tumour  in  a  case  of 
quarter-evil  were  not  materially  affected.  The  possibility  of  those 
calves  which  possess  a  natural  immunity  being  classed  as  protected 
by  the  inoculation  must  be  admitted,  and  the  efficacy  and  safety 
of  the  process  is  by  no  means  established. 

MALIGNANT  CEDEMA. 

The  disease  known  by  surgeons  as  progressive  gangrene,  gan- 
grenous emphysema,  or  surgical  gangrene,  has  been  shown  by  the 
researches  of  Chauveau,  Arloing,  Rosenbach  and  Babes,  to  be 
due  to  a  bacillus  identical  with  the  microbe  septique  of  Pasteur  and 
the  bacillus  of  malignant  oedema  of  Koch.  The  bacillus  or  its  spores 
may  be  spread  by  the  neglect  of  antiseptics.  The  disease  occurs 
especially  after  compound  fractures  and  gun-shot  wounds. 

If  a  guinea-pig  is  subcutaneously  inoculated  with  earth,  putrid 
fluid,  or  hay  dust,  death  frequently  occurs  in  from  twenty-four  to 
forty- eight  hours.  At  the  autopsy  the  most  characteristic  symptom 
is  a  widespread  subcutaneous  oedema  accompanied  by  air-bubbles. 
This  originates  from  the  point  of  inoculation,  and  contains  a 
clear  reddish  liquid  full  of  motile  and  n on- motile  bacilli.  The 
internal  organs  are  little  changed,  the  spleen  is  enlarged  and  of  a 
dark  colour,  and  the  lungs  are  hypersemic,  and  have  hsemorrhagic 
spots.  Examined  immediately  after  death,  few  or  no  bacilli  are 
detected  in  the  blood  of  the  heart,  but  in  that  of  the  spleen,  liver, 
lungs,  and  other  organs,  in  the  peritoneal  exudation,  and  in  and 
upon  the  serous  coating  of  the  abdominal  organs,  they  are  present  in 
large  numbers.  If,  on  the  other  hand,  the  animal  is  not  examined 
until  some  time  after  death,  the  bacilli  are  found  in  the  blood  of 
the  heart,  and  distributed  all  over  the  body. 

Bacillus  (Edematis  Maligni,  Koch  (Pasteur's  Septicaemia). — 


MALIGNANT    (EDEMA. 


221 


Rods  from  3  to  3'5  /*,  long  and  1  to  1*1  /x  wide  ;  they  mostly  lie 
in  pairs,  and  then  appear  to  be  double  this  length.  The  rods  are 
rounded  at  their  ends,  and  form  threads  which  are  sometimes  straight, 
but  more  commonly  curved.  In  stained  preparations  they  have  a 
somewhat  granular  appearance.  They  are  motile,  possessing  flagella, 
and  form  spores.  The  bacilli  are  distinguished  from  anthrax  bacilli 
by  their  being  somewhat  thinner,  by  their  rounded  ends,  and  by  their 
motility.  Moreover,  anthrax  bacilli  never  appear  as  threads  in  fresh 
blood,  and  are  differently  distributed  throughout  the  body.  They 
nre  anaerobic,  and  can  be  cultivated  on  blood  serum  and  on  neutral 
solution  of  Liebig's  meat  extract  in  an  atmosphere  of  carbonic  acid. 
By  embedding  material  containing  bacilli  in  nutrient  agar-agar 


\ 


FIG.  103.     BACILLI  OF  MALIGNANT  (EDEMA  x   950.     From  the  subcutaneous  tissue 
of  a  guinea-pig.   (BAUMGARTEN.) 

and  nutrient  gelatine,  characteristic  cultivations  are  obtained.  The 
following  process  may  be  adopted  to  obtain  a  pure  cultivation.  A 
mouse  inoculated  subcutaneously  with  dust,  as  a  rule,  dies  in  one 
to  two  days.  It  is  then  pinned  out,  back  uppermost,  011  a  slab  of 
wood,  and  the  hair  singed  with  a  Paquelin's  cautery  from  one  hind 
leg  up  to  the  neck,  across  the  latter,  and  down  again  to  the  opposite 
hind  leg.  Following  the  cauterised  line,  the  skin  is  cut  through  with 
.sterilised  scissors,  and  the  flap  turned  back  and  pinned  out  of  the 
way.  With  curved  scissors  little  pieces  of  the  subcutaneous 
cedematous  tissue,  in  the  neighbourhood  of  the  inoculated  spot,  are 
cut  out,  and  sunk  with  a  platinum  needle  in  a  1  per  cent,  nutrient 
agar-agar,  or  5  per  cent,  nutrient  gelatine.  Fragments  of  tissue  may 
also  be  embedded  by  the  method  already  described  for  anaerobic 
bacteria. 


222 


INFECTIVE    DISEASES. 


The  inoculated  tubes  are  placed  in  the  incubator.  In  a  few  hours 
a  whitish  turbidity  spreads  out  from  the  piece  of  tissue,  and  upwards 
in  the  needle  track.  Examined  microscopically,  the  turbidity  is  found 
to  be  due  solely  to  the  development  of  the  bacilli  of  oedema.  The 
surface  exposed  to  the  air  exhibits  no  trace  of  the  bacilli.  To 
investigate  the  tubes  microscopically,  a  sterilised  glass  tube  with  a 
capillary  end  may  be  used,  with  its  neck 
plugged  with  sterilised  cotton -wool,  and 
provided  at  the  mouth  with  a  suction  ball. 
The  capillary  end  is  thrust  into  the  cultiva- 
tion, and  a  small  fragment  removed  by 
aspiration. 

In  the  course  of  the  first  day  the  bacilli 
spread  throughout  a  great  part  of  the 
agar-agar  in  such  a  way  that  a  more  or  less 
equally  diffused  cloudiness  of  the  medium  en- 
sues, with  subsequent  appearance  of  strongly 
marked  clouds  or  lines  of  turbidity.  At  the 
same  time  gas- bubbles  develop  along  the 
needle  track,  and  a  collection  of  liquid  takes 
place,  while  spore-formation  also  commences. 
The  following  day  these  appearances  are  more 
marked,  the  opacity  is  more  pronounced,  the 
development  of  gas  increases,  and  the  liquid 
contains  more  spore-forming  bacilli  and  nu- 
merous free-spores. 

The  nutrient -gelatine  cultures  during  the 
first  day  show  no  macroscopic  change,  but 

FIG.   104.     "PURE-CULTURE   after  a  few  days  the  piece  of  tissue  is  sur- 
OF  BACILLUS  OF  MALIG-  rounded  with  a  white  halo.     This  gradually 
spreads  in  all  directions,  and  is  apparently 
beset  with  hairs.     The  gelatine  liquefies,  and 
the  fragment  of  tissue,  degenerated  bacilli,. 

and  spores,  sink  to  the  bottom.  The  cultivation  is  also  very 
characteristic  in  a  ^  per  cent,  nutrient  agar-agar.  If  placed  in 
the  incubator,  in  a  few  hours  a  cloudiness  forms  around  the  piece  of 
embedded  tissue,  which  is  caused  by  bacilli  gradually  spreading  in  all 
directions  in  the  nutrient  medium.  Mice  inoculated  from  these 
cultivations  die  more  quickly  than  from  the  original  infection  from 
dust.  On  potatoes  they  are  cultivated  by  introducing  a  piece  of  liver 
or  other  tissue  containing  the  bacilli,  into  the  interior  of  a  sterilised 
potato,  and  incubated  at  38°  C.  The  bacillus  is  not  deprived  of  its 
virulence  by  cultivation. 


NANT  (EDEMA  IN  GRAPE- 
SUGAR  GELATINE  (FRAN- 
KEL  and  PFEIFFER). 


j 
/ 


MALIGNANT    (EDEMA.  223 

The  spores  of  the  oedema  -bacilli  appear  to  be  very  widely  dis- 
tributed. They  are  found  in  the  upper  cultivated  layers  of  the  soil, 
in  hay  dust,  in  decomposing  liquids,  and  especially  in  the  bodies  of 
suffocated  animals,  which  are  left  to  decompose  at  a  high  temperature. 
From  any  of  these  sources  animals  can  be  successfully  inoculated. 
The  bacillus  is  not  only  pathogenic  in  guinea-pigs,  rabbits,  and 
mice,  but  also  in  man  and  in  farm  animals,  including  calves  but  not 
cattle.  Pure  -cultures  inoculated  in  animals  produce  oedema  at  the 
seat  of  inoculation 
without  appreciable 
gas  formation  and 
without  any  putrefac- 

tive odour.    The  odour  * 

and     frothy     effusion 
resulting  from  the  in- 

oculation  of  earth  are      /  ^  .^ 

due  to  other  bacteria,  \i 

which   are    introduced  f 

simultaneously      with       X*~  /     4^ 

the    bacilli    of    malig-  \  ^  V 

iiant     cedema.        The  _^   •     ^^^ 

spleen     is     sometimes 
slightly  enlarged.     By 
touching  with  a  cover- 
glass    the    capsule    of  FlG>  m    BACILLI  OF  MALIGNANT  (EDEMA  x  1000. 
the  spleen,  or   by  ex-       From  an  agar  culture  (FBANKEL  and  PFEIFFER). 
amining     the     serous 

effusion,  the  bacilli  are  found  in  abundance  ;  but  if  a  preparation 
is  made  from  the  interior  of  the  spleen  or  from  the  blood  of 
the  heart,  no  bacilli  will  be  found  until  several  hours  after 
death.  In  this  respect  there  is  a  marked  difference  from 
anthrax.  Another  difference  is  shown  in  spore-formation,  which 
occurs  in  the  living  body  in  malignant  oedema,  but  never  in 
anthrax.  Animals  which  recover  from  the  disease  are  said  to 
be  protected. 

Protective  Inoculation.  —  Roux  and  Chamberland  produced 
immunity  by  injecting  the  chemical  products  in  the  nitrate  obtained 
from  cultures  in  broth.  The  serum  from  fatal  cases  will,  it  is  said, 
confer  immunity  on  other  animals. 

There  is  a  variety  of  this  bacillus  in  soil  according  to  Fliigge, 
agreeing  in  morphological  and  cultural  but  not  in  pathogenic 
characters. 


224  INFECTIVE   DISEASES. 

RAG-PICKERS'  SEPTICAEMIA. 

Hag-pickers'  disease  has  a  resemblance  to  anthrax  or  wool-sorters' 
disease.  After  death  the  spleen  is  found  to  be  enlarged,  the  in- 
ternal organs  are  congested,  and  there  are  haemorrhages  on  the 
serous  membranes.  Bordoni -Uffreduzzi  isolated  bacilli  which  are 
quite  easily  distinguished  from  anthrax  bacilli.  They  were  found 
in  the  blood  and  in  sections  of  the  internal  organs. 

Proteus  hominis  eapsulatus. — Rods  with  rounded  ends,  singly, 
in  pairs,  and  in  filaments,  somewhat  smaller  than  anthrax  bacilli, 
and  often  irregular  in  form.  Spore-formation  not  described  ;  they 
have  a  well-marked  capsule.  Colonies  are  circular,  appearing  at 
first  granular,  and  later  possessing  a  filamentous  structure.  In 
the  depth  of  gelatine  they  grow  in  the  shape  of  a  round-headed 
nail,  like  a  culture  of  Friedlander's  pneumococcus.  On  the  surface  of 
gelatine  they  form  a  shining  white  layer.  On  agar  the  growth  is 
somewhat  transparent.  On  potato  a  moist,  glistening  film  gradually 
spreads  over  the  surface.  They  do  not  liquefy  blood  serum,  and 
the  growth  is  similar  to  that  obtained  on  agar.  They  prove  fatal 
to  mice  and  dogs,  but  rabbits  and  guinea-pigs  are  not  very  sus- 
ceptible. Dogs  die  usually  on  the  second  day  after  intravenous 
injection,  and  after  death  there  is  congestion  of  the  internal  organs 
and  of  the  intestinal  mucous  membrane.  (Edema  is  produced  at 
the  seat  of  inoculation  in  mice.  There  are  haemorrhages  in  the 
lymphatic  glands,  and  congestion  of  the  liver  and  kidneys.  Similar 
organisms  have  been  described  by  Kolb  and  by  Babes  in  purpura 
hsemorrhagica. 

SEPTICAEMIA  OF  GUINEA-PIGS. 

Guinea-pigs  and  mice  sometimes  die  of  septicaemia,  characterised 
by  congestion  of  the  lungs,  liver,  and  kidneys,  inflamed  peritoneum, 
pleural  and  pericardial  exudation,  congested  spleen,  and  congestion 
of  the  mucous  and  serous  coats  of  the  intestine.  Klein  isolated 
a  bacillus  from  the  blood  and  the  internal  organs  in  these  cases. 

Bacillus  of  Septicaemia  in  Guinea-pigs. — Rods  with  rounded 
ends,  motile,  with  pleomorphic  forms,  cocci,  short  rods  and  filaments. 
Colonies  appear  as  small,  circular,  white  dots,  which  enlarge  and 
become  irregular  in  outline.  In  the  depth  of  gelatine  a  white 
filament  develops,  and  on  the  surface  the  growth  rapidly  spreads 
with  a  crenated  outline.  Broth  becomes  turbid,  and  after  the  second 
day  a  copious  white  sediment  is  deposited.  Spore-formation  not 
observed. 


DESCRIPTION    OF    PLATE    VI. 
Bacillus  Murisepticus, 

FIG.  1. — From  a  section  of  a  kidney  of  a  mouse  which  had  died  after  inocula- 
tion with  a  pure-cultivation  of  the  bacillus.  With  moderate  amplification, 
the  white  blood-corpuscles  have  a  granular  appearance,  and  irregular 
granular  masses  are  scattered  between  the  kidney  tubules.  Stained  by 
Gram's  method  with  eosin.  x  200. 

FlG.  2. — Part  of  the  same  preparation  with  high  amplification.  The  granular 
appearances  are  found  to  be  due  to  the  presence  of  great  numbers  of 
extremely  minute  bacilli,  x  1500. 


Plate  VI 


>-  > 


i 

< 


. 


i. 


i 


Ce 


Figl. 


V, 


> 
I 


Fig,  2. 


BACILLUS     MURISEPTICUS 


SEPTICAEMIA   OF  MICE.  225 

According  to  Wooldridge,  the  chemical  products  of  this  bacillus, 
separated  by  filtration,  produce  on  inoculation  immunity  against 
virulent  bacilli. 

SEPTICAEMIA  OF  MICE. 

Mice  inoculated  with  a  minimum  quantity  of  putrid  fluid  often 
die  of  septicaemia.  They  rapidly  sicken,  their 
eyes  inflame,  their  eyelids  stick  together,  they 
become  soporific,  and  death  occurs  in  forty  to 
sixty  hours.  There  is  slight  oedema  at  the  seat 
of  inoculation,  and  enlargement  of  the  spleen  ; 
the  bacilli  are  found  free  and  in  the  interior 
of  white  corpuscles,  both  in  the  cedematous 
tissue  and  in  the  blood  capillaries. 

Bacillus  of  Septicaemia  of  Mice  (Koch). 
—Extremely  minute  bacilli,  -8  to  1  p  long,  and 
•1  to  -2  fj.  broad,  and  filaments.  In  cultivations 
in  gelatine  they  do  not  appear  to  make  threads, 
but  the  bacilli  lie  together  in  masses.  Spores 
have  been  observed.  The  bacilli  are  probably 
non-motile.  They  are  most  commonly  in  the 
interior  of  white  blood  corpuscles.  In  these 
they  increase,  and  in  many  cases  a  white  blood 
cell  is  represented  only  by  a  mass  of  bacilli. 

A  minimal  quantity  of  blood  containing  the 
bacilli  produces  the  disease  if  inoculated  in 
house-mice  or  sparrows.  Field-mice  have  an 
immunity.  Rabbits  and  guinea-pigs  inoculated 

in  the  ear  suffer  only  from  a  local  erythema,   , 

J  J  '    FIG.   106.—  PURE  CUL- 

which   disappears   after  five  or  six   days,   and       TIVATION     OF     THE 

renders   them    for    a    time   immune.     Rabbits      BACILLUS  OF  SEPTI- 
inoculated  in  the  cornea  suffer  from  an  intense      C^MIA   OF   MICE   IN 


.    a  ,.          »  ,,  mi      i_     -IT  .e          •          NUTRIENT  GELATINE. 

inflammation  of  the  eyes.    The  bacilli  form  m 

J  After  two  days. 

plate-cultivations  scarcely  perceptible  cloud-like 

specks,  and  in  a  test-tube  of  nutrient  gelatine  they  form  a  delicately 
clouded  cultivation  along  the  needle  track. 

An  identical  bacillus  has  been  isolated  in  swine  measles. 


15 


(CHAPTER  XVT. 

SEPTICEMIA       OF       BUFFALOES. SEPTIC       PLEURO-PNEUMONIA       OF 

CALVES. — SWINE  FEVER.—  SEPTICAEMIA  OF  DEER.— SEPTICAEMIA 
OF  RABBITS.  —  FOWL  CHOLERA.  —  FOWL  ENTERITIS.  —  DUCK 
CHOLERA. — GROUSE  DISEASE. 

THERE  are  several  varieties  of  septicaemia  occurring  naturally  in 
buffaloes,  deer,  calves,  and  birds,  and  artificially  induced  by  inocula- 
tion of  rabbits  with  septic  material.  They  are  associated  with 
bacteria  which  agree  in  their  morphological  and  cultural  characters, 
though  in  some  cases  differing  in  their  pathogenic  properties.  As 
the  differences  between  the  bacteria  cultivated  from  these  different 
sources  is  not  greater  than  the  differences  which  exist  between 
the  morphological,  biological,  and  pathogenic  effects  of  varieties  of 
the  tubercle  bacillus,  it  will  be  convenient  and  fully  justifiable  to 
follow  Hueppe  and  Baumgarten,  and  regard  them  as  varieties  of 
the  bacillus  of  hcemorrhagic  septicaemia. 

EPIDEMIC  DISEASE  OF  BUFFALOES. 

Oreste  and  Armanni  investigated  an  epidemic  among  herds  of 
young  buffaloes  in  Italy  (Eilffd-seuche).  The  disease  was  extremely 
acute,  death  occurring  in  from  twelve  to  twenty -four  hours.  It  was 
probably  identical  with  an  epidemic  disease  described  by  Bellinger  in 
deer.  The  symptoms  were  fever,  rapid  pulse,  discharge  of  mucus 
from  the  nose  and  mouth,  and  a  local  swelling  of  the  head  and  face 
leading  to  suffocation.  The  only  marked  feature  after  death  was 
hsemorrhagic  inflammation  of  the  small  intestine. 

The  bacilli  were  identical  with  those  found  by  Schiitz  in  swine 
fever.  Cultures  inoculated  in  young  buffaloes  produced  the  disease. 
The  bacilli  were  pathogenic  to  mice,  guinea-pigs,  rabbits,  pigeons, 
and  fowls,  death  taking  place  in  from  one  to  three  days. 

226 


EPIDEMIC   DISEASE   OF   DEER   AND    BOARS.  227 

SEPTIC  PLEURO-PNEUMONIA  IN  CALVES. 

Septic  pleuro-pneumoriia  is  a  disease  which  attacks  young  calves 
within  the  first  two  months  after  their  birth.  Percussion  and 
auscultation  reveal  lung  mischief.  The  disease  is  very  rapid  and 
fatal,  death  occurring  on  the  second  or  third  day.  In  the  less 
acute  cases  one  or  more  lobes  of  the  lungs  are  found  after  death 
in  a  state  of  lobular  and  inter-lobular  pneumonia.  The  inter-lobular 
connective  tissue  is  distended  with  exudation,  giving  rise  to  white 
or  yellowish  bands  between  the  inflamed  lobules,  which  produce  a 
marbled  appearance,  recalling  the  condition  of  the  lungs  in  infectious 
pleuro-pneumonia.  The  internal  organs  are  congested,  and  there 
are  very  often  haemorrhagic  spots  on  the  mucous  and  serous  coats 
of  the  small  intestine.  All  the  organs  contain  rods  identical  with 
those  of  septicaemia  of  rabbits.  Rabbits,  guinea-pigs,  and  mice  were 
infected.  A  calf  was  injected  in  the  pleural  cavity  with  a  broth - 
culture,  and  died  in  twenty  hours. 

SWINE  FEVER. 

This  disease  will  be  described  in  a  separate  chapter.  Several 
bacteria  have  been  isolated  by  different  investigators.  In  swine 
fever  in  Germany  (Schwein-seuche)  Loffler  and  Schiitz  isolated  a 
bacillus  which  has  been  identified  with  the  bacillus  isolated  by 
Salmon  and  Smith  from  hog-cholera  in  America,  and  with  the 
bacillus  of  rabbit  septicaemia  and  of  fowl  cholera. 

EPIDEMIC  DISEASE  OF  DEER  AND  BOARS. 

A  very  fatal  epizootic  (Wildseuche)  occurred  in  the  royal  game 
preserves  near  Munich,  destroying  one  hundred  and  fifty-three  deer 
and  two  hundred  and  thirty-four  boars  (Bellinger).  The  disease 
lasted  from  twelve  hours  to  six  days.  In  the  less  acute  cases  pneu- 
monia and  pericarditis  supervened.  In  cattle  there  was  also  severe 
haemorrhagic  inflammation  of  the  small  intestine.  In  another  form 
it  produced  swelling  of  the  head,  face,  neck,  and  tongue.  The 
virus  proved  fatal  to  rabbits  in  six  to  eight  hours,  and  to  sheep  and 
goats  in  about  thirty  hours.  A  pig  inoculated  with  a  few  drops  of 
blood  died  in  twenty-two  hours.  Kitt  also  investigated  this  malady. 
The  bacteria  were  found  to  be  identical,  in  their  appearance  and 
pathogenic  properties,  with  extremely  virulent  bacteria  from  swine 
fever.  Schiitz  distinguished  them  from  the  bacteria  obtained  from 
swine  fever  by  their  pathogenic  effect  on  pigeons,  but  cultures 
obtained  from  swine  fever  do  not  act  uniformly  in  this  respect. 


228  INFECTIVE    DISEASES. 

SEPTICAEMIA  IN  RABBITS. 

Koch  minutely  investigated  a  disease  of  rabbits  produced 
by  inoculation  with  impure  river  water  and  with  putrid  meat 
infusion.  Bacteria  are  found  in  the  blood  in  abundance,  and  may 
be  readily  cultivated. 

The  smallest  quantity  inoculated  subcutaneously  or  in  the  cornea 
of  a  rabbit  produces  a  rise  of  tempera- 
ture and  laboured  breathing  after  ten 
to  twelve  hours,  and  death  in  sixteen 

^^         ^  f        ^ 

to    twenty    hours.      The    spleen    and 
lymphatic    glands    are    found     to    be 

FIG.  107. -BACTERIUM  OF  RABBIT  enlarged,    and    the    lungs    congested, 
SEPTICAEMIA;  BLOOD  OF  SPAR-  °  .  ' 

ROW,  x  700  (Kocn).  Dut  there  are  no  extravasations,  and  no 

peritonitis.      Mice  and  birds  are  very 
susceptible ;    guinea-pigs  and  white  rats  have  an  immunity. 

DAVAINE'S  SEPTICAEMIA. 

A  disease  was  produced  by  Davaine  by  injecting  rabbits  with 
putrid  blood.  Rabbits,  mice,  fowls,  pigeons,  and  sparrows  are  sus- 
ceptible, and  guinea-pigs  and  rats  are  insusceptible  to  the  bacteria 
found  in  this  disease.  Rabbits  inoculated  with  a  trace  of  blood  con- 
taining the  bacteria,  or  with  a  culture,  died  in  from  twenty-four  to 
thirty-six  hours.  The  spleen,  liver,  lungs,  and  intestines  are  highly 
congested,  and  sometimes  extravasations  and  peritonitis  are  found. 

FOWL  CHOLERA. 

Fowl  cholera  is  an  epidemic  disease  of  the  poultry-yard  much 
dreaded  in  France,  and  well  known  through  the  researches  of 
Perroncito,  Toussaint,  Pasteur,  and  Kitt. 


•*• 


,-,  FIG.  109.  —  BACTERIUM    OF  FOWL 

1  ™    ^  BAC™Mf°F  F7L  °*OL1f  *»  CHOLERA,  x  2500.  Muscle  juice 

x  1200.    From  blood  of  inoculated  Fowl. 


Fowls  suffering  from  the  disease  usually  die  in  from  twenty-four 
to  forty-eight  hours.     The  disease  shows  itself  by  the  fowls  becoming 


FOWL    CHOLERA.  229 

somnolent.  They  suffer  from  weakness  of  the  legs,  and  their  wings 
trail.  There  is  frequently  diarrho3a,  with  slimy  greenish  evacuations, 
and  death  usually  ensues  after  a  slight  convulsive  attack.  On 
making  a  post-mortem  examination  the  viscera  will  be  found  to  be 
congested,  and  there  is  intense  inflammation  of  the  mucous  membrane 
of  the  intestine,  with  haemorrhages. 

The  blood  from  the  heart,  and  the  intestinal  contents,  contain 
the  bacilli  which  were  at  one  time  believed  to  be  peculiar  to 
this  disease.  Inoculation  subcutaneously,  or  administration  with 
food,  of  a  small  quantity  of  a  broth  cultivation  will  produce  death 
in  twenty-four  to  thirty-six  hours.  Pigeons,  pheasants,  sparrows, 


fn      ^fcf^3;^i 

^ffs?''^" 


FIG.  110. — BACTERIUM  OF  FOWL  CHOLERA.    Section  from  liver  of  Fowl   x  700 

(FLUGGE). 

rabbits,  and  mice  are  susceptible.  In  guinea-pigs,  sheep,  and  horses, 
an  abscess  develops  at  the  seat  of  inoculation.  Rabbits  are  readily 
infected  by  sprinkling  a  broth-cultivation  on  cabbage  leaves  or  any 
suitable  food.  It  was  with  this  microbe  that  Pasteur  proposed  to 
eradicate  the  plague  of  rabbits  in  Australia. 

Fowl  cholera  has  an  additional  interest,  as  it  was  with  this 
disease  that  Pasteur  first  investigated  the  attenuation  of  virus. 
Broth -cultures  which  were  several  months  old  were  found,  when 
injected,  to  produce  apparently  only  a  local  effect.  This  weakening 
of  the  virus  was  attributed  by  Pasteur  to  exposure  to  oxygen. 
After  recovery  the  fowls  were  protected  against  the  action  of 
virulent  cultures,  while  fowls  not  immunised  died  the  following  day. 
Kitt,  by  working  with  pure-cultures  on  solid  media,  showed  that  the 


X^TBRATT?^ 

[  "     OF   THK  *N 

I  UNIVERSITY  1 


230  INFECTIVE    DISEASES. 

weakening  was  not  due  to  prolonged  exposure  to  oxygen,  but  that 
old  contaminated  broth- cultures  after  a  time  completely  lost  their 
power,  owing  to  the  antagonism  of  the  bacteria  accidentally  present. 
Filtered  broth- cultures  contain  the  toxic  products  of  the  bacillus, 
and  produce  slight  illness  and  subsequent  immunity. 

FOWL  ENTERITIS. 

Fowl  enteritis  is  an  acute  infectious  disease  of  fowls,  the  course 
and  symptoms  of  which  are  regarded  by  Klein  as  distinct  from  fowl 
cholera.  The  fowls  suffer  from  diarrhoea,  with  liquid  greenish 
evacuations,  but  are  never  somnolent,  and  death  occurs  in  one  or 
two  days.  After  death  the  mucous  membrane  of  the  intestine  is 
found  to  be  congested,  and  coated  with  grey  or  yellowish  mucus; 
the  liver  is  congested,  spleen  enlarged,  and  lungs  normal.  There  are 
a  few  bacilli  in  the  blood  of  the  heart,  very  many  in  the  spleen  and 
liver,  and  they  are  in  the  form  of  a  pure-culture  in  the  mucus  of 
the  intestine.  Klein  says  that  the  bacilli  are  a  little  longer  and 
thicker  than  those  found  in  fowl  cholera,  which  they  only  slightly 
resemble,  and  that  the  course  of  the  disease,  the  symptoms  and 
pathological  appearances,  definitely  distinguish  it  from  fowl  cholera, 
but  that  nevertheless  it  belongs  to  the  same  family  of  bacilli 
Pigeons  are  said  to  be  insusceptible,  rabbits  only  slightly  susceptible. 
By  feeding  and  by  subcutaneous  inoculation  the  disease  can  be 
communicated  to  healthy  fowls,  but  there  is  no  sign  of  illness  until 
the  fourth  day.  As  regards  attenuation,  the  bacilli  behave  like 
those  from  cases  of  fowl  cholera. 

DUCK  CHOLERA. 

Duck  cholera  is  an  epidemic  disease  of  ducks  which  was  investi- 
gated by  Cornil.  The  symptoms  are  similar  to  those  of  fowl  cholera. 
They  suffer  from  diarrhoea  and  weakness,  followed  by  death  in  two 
or  three  days. 

The  bacillus  cultivated  from  the  blood  of  ducks  is  pathogenic  in 
ducks  but  not  in  fowls  or  pigeons,  and  large  doses  are  required  to 
kill  rabbits. 

GROUSE  DISEASE. 

Grouse  disease  is  an  acute  infectious  disease  of  red  grouse. 
According  to  Klein  the  chief  pathological  feature  is  severe  pneu- 
monia ;  there  is  also  patchy  redness  of  the  serous  and  mucous  linings 


GROUSE    DISEASE. 


• 


-Y  O 

juv 


FIG.  111.— BACILLUS  OF  H^MORRHAGIC 
SEPTICAEMIA.  Blood  of  a  Rabbit  after 
death  from  Septicaemia  x  950  (BAUM- 
GARTEN). 


of  the  intestine,  and  the  liver  is  congested  and  dark,  but  the  spleen 
is  not  enlarged.     The  bacilli  are  found  in  the  heart,  lungs,  and  liver, 
and  in  the  extravasated  blood. 
Cultures  inoculated  in  mice  and 
guinea-pigs  produce  pneumonia 
and  death.      Sparrows  are  sus- 
ceptible, and  other  small  birds. 
Fowls,  pigeons,  and  rabbits  are 
insusceptible. 

Bacillus  of  Haemor- 
rhagic  Septicaemia. — Very 
short  rods,  with  rounded  ends, 
*6  to  '7  JJL  in  width  and  1'4  //.  in 

length.  In  stained  preparations  the  rods 
are  observed  to  be  deeply  stained  at  the 
ends  and  to  have  a  clear  interval  in  the 
middle;  they  were  on  this  account  mis- 
taken by  earlier  observers  for  dumb-bell 
micrococci  or  diplococci.  They  are  non- 
motile,  and  spore-formation  is  unknown. 
They  grow  readily  in  the  ordinary  media. 
The  colonies  in  nutrient  gelatine  appear 
about  the  third  day.  They  are  circular 
in  form,  with  a  sharp  dark  outline,  and 
of  a  yellow  colour,  lighter  at  the  peri- 
phery. Later,  the  central  zone  is  finely 
granular,  and  of  a  dark  yellowish-brown 
colour,  with  the  lighter  peripheral  zone 
more  clearly  defined.  In  the  depth  of 
gelatine  a  delicate  filament  develops  in 
the  track  of  the  needle,  composed  of 
minute  spherical  colonies,  somewhat  trans- 
parent, and  yellowish-white  in  colour. 
At  the  point  of  puncture  there  may  be  no 
growth  visible,  or  a  flat  and  very  limited 
growth.  Inoculated  on  the  surface  of 
nutrient  media  a  thin  layer  develops, 
with  an  irregular  serrated  and  thickened 
border.  On  potato  different  results  have 
been  obtained  by  different  observers.  Some  maintain  that  a 
greyish-white  or  yellowish  film  will  develop  at  the  temperature  of 
the'blood  ;  but  according  to  Caneva,  the  bacilli,  whatever  their  source, 


FIG.     112.  —  BACILLUS    OF 

H.EMORRHAGIC   SEPTICAEMIA 

(Rabbit  Septicaemia).  Pure- 
culture  in  Gelatine  after 
four  days  (B AUMG ARTEN.  ) 


232  INFECTIVE    DISEASES. 

will  not  grow  on  potato,  while  Bunzl-Feclern  maintains  that  the 
bacilli  from  fowl  cholera  and  rahbit  septicaemia  do  grow  upon  potato, 
but  those  from  septicaemia  in  deer,  buffaloes,  and  swine  do  not. 
Opinions  differ  with  regard  to  their  action  on  milk.  The  reaction 
for  phenol  and  indol  is  given  in  all  cases,  except  with  cultures 
obtained  from  septicaemia  of  buffaloes.  The  virulence  of  the  bacilli 
may  be  diminished  and  attenuated,  but  it  may  subsequently  be 
restored  by  successive  inoculation  in  animals.  The  pathological 
lesions  vary  in  different  animals.  The  most  common  result  is  con- 
gestion of  the  internal  organs  and  haemorrhage.  The  bacilli  culti- 
vated from  cattle  or  deer  produce  fatal  results  when  inoculated 
in  swine.  The  bacilli  from  any  of  these  sources  inoculated  in 
pigeons  will  produce  fowl  cholera,  but  the  bacilli  isolated  by  Schiitz 
from  swine,  and  those  from  deer,  are  not  fatal  to  fowls.  Further, 
the  bacilli  cultivated  from  swine  fever  are  fatal  to  guinea-pigs, 
while  the  bacilli  from  rabbit  septicaemia  have  very  little  effect  upon 
them.  The  bacilli  have  been  found  in  association  with  diseases  of 
cattle,  swine,  deer,  birds,  rabbits,  and  mice,  and  have  been  cultivated 
from  healthy  mucous  membrane.  Yeranus  Moore  found  the  bacilli 
in  the  mucus  from  the  upper  air  passages,  of  71  per  cent,  of  cattle, 
85  per  cent,  of  cats,  and  33  per  cent,  of  dogs.  From  these  sources 
inoculations  were  made  in  rabbits,  and  rapidly  fatal  septicaemia 
was  produced,  associated  in  less  acute  cases  with  peritonitis,  pleurisy, 
and  pericarditis. 


CHAPTER   XVII. 

PNEUMONIA. — INFECTIOUS   PLEURO-PNEUMONIA    OF    CATTLE. — 
INFLUENZA. 

ACUTE  CROUPOUS  PNEUMONIA. 

PNEUMONIA  is  an  acute  inflammation  of  the  lungs  with  fibrinous 
infiltration  of  the  air  vesicles  and  interstitial  tissue.  There  are 
varieties  of  pneumonia,  and  one  form  is  commonly  believed  to  be 
infectious. 

The  lung  passes  through  three  stages — engorgement,  red  hepati- 
sation,  and  grey  hepatisation.  In  the  first  stage  the  lung  is  of 
a  deep  red  colour,  but  still  vesicular ;  in  the  second  stage  the 
affected  part  is  more  or  less  solid,  and  has  the  consistency  of  liver, 
owing  to  the  fibrinous  lymph  which  is  poured  out  into  the  alveolar 
cavities.  In  the  grey  hepatisation,  the  exudation  contains  more 
leucocytes  and  less  fibrin,  and  this  is  followed  by  the  stage  of 
suppurative  softening  and  final  absorption.  The  sputum  at  the 
commencement  of  the  disease  is  rusty,  from  the  presence  of  blood, 
and  later  on  has  the  appearance  of  prune  juice.  Examination  of 
the  sputum  by  Gram's  method  will  reveal  numerous  micro-organisms, 
and  two  of  these  are  deserving  of  special  study — the  pneumococcus 
of  Friedlander,  which  is  present  in  a  considerable  proportion  of 
cases,  and  Sternberg's  micrococcus,  which  was  found  in  sputum  by 
Talamon. 

In  1888,  there  was  considerable  prevalence  of  pneumonia  in 
Middlesbrough,  with  strong  tendency  to  occur  in  groups  of  cases; 
but  there  was  admittedly  room  for  doubt  whether  the  clinical 
and  post-mortem  appearances  were  not  identical  with  ordinary 
pneumonia.  Dr.  Ballard  maintained  that  there  were  facts  and 
considerations  which  appeared  to  show  that  the  disease  was  com- 
municable from  the  sick  to  the  healthy,  and  that  it  was  a  specific 
febrile  disease,  and  Klein  isolated  and  described  the  micrococcus 
present  in  these  cases. 

Bacterium    Pneumonias    Crouposae    (Pneumococcus,  Fried- 


234 


INFECTIVE    DISEASES. 


FIG.  113.— BACTERIUM  PNEUMONLE 
CROUPOS^E,  FROM  PLEURAL  CAVITY 
OF  A  MOUSE,  x  1500.  A,  B. 
Thread-forms.  C,  D,  E.  Short 
rod-forms.  G.  Diplococci.  H. 
Cocci.  I.  Streptococci.  (Zopf.) 


lander). — Cocci  ellipsoidal  and  round,  singly,  or  in  pairs  (diplococci), 

rods  and  thread-forms.  The  cell- 
membrane  thickens,  and  develops 
into  a  gelatinous  capsule,  which  is 

(round  if  the  coccus  is  single,  and 
ellipsoidal  if  the  cocci  occur  in  pairs 
or  in  rod-forms.  Cultivated  in  a 
test-tube  of  nutrient  gelatine  they 
grow  in  the  form  of  a  round- 
headed  nail,  without  liquefaction 
of  the  gelatine  (Fig.  114).  The 
cocci  when  artificially  cultivated 
have  no  capsule,  but  it  again 
appears  after  their  injection  into 
animals, 
The  cocci 
can  also  be 
cultivated 

on  blood  serum  and  on  boiled  potatoes. 
They  occur  in  pneumonic  exudation.  In- 
oculation of  dogs  with  a  cultivation  of  the 
cocci  occasionally  gave  positive  results;  but 
in  rabbits  no  results  followed.  Guinea- 
pigs  proved  to  be  susceptible  in  some  cases ; 
but  thirty- two  mice,  after  injection  of  a 
cultivation  diffused  in  sterilised  water,  into 
the  lungs,  died  without  exception.  The 
lungs  were  red  and  solid,  and  contained  the 
cocci,  which  were  also  present  in  the  blood, 
and  in  enormous  numbers  in  the  pleura! 
exudation.  Inhalation  experiments  by  spray- 
ing the  cocci  diffused  in  water  into  mouse 
cages  produced  pneumonia  and  pleurisy  in 
three  out  of  ten  mice. 

The  nail-shaped  cultivation  is  not  always 

produced,  nor  are  these  conclusions  accepted 

—  -  IIG.  114. — FBIEDLANDBBS 


by  all  investigators. 

METHODS   OF  STAINING    FRIEDLANDER's 
PNEUMOCOCCUS. 


PNEUMOCOCCUS.  Pure- 
culture  in  nutrient- 
gelatine  four  days  old 
(BAUMGARTEN). 


Cover-glass  preparations  of  pneumonic  sputum  or  exudation  may  be 
treated  as  follows  :— 


PNEUMONIA. 


235 


(a) 

(b) 

brown. 
balsam. 

(c) 
hours  ; 

(<?) 
out. 


Stain  by  the  method  of  Gram,  and  after-stain  with  eosin. 
Treat  with  ace  tic  acid,  then  stain  with  gentian-violet  or  Bismarck- 
Examine  in   distilled  water,   or  dry  and  preserve  in   Canada 

Float  them  on  weak  solutions  of  the   aniline  dyes  twenty-four 
differentiation  between  coccus  and  capsule  is  thus  obtained. 
Stain  with  osmic  acid  ;  the  contour  of  the  capsules  is  brought 


; 


FIG.  115. — CAPSULE-COCCI  FROM  PNEUMONIA,   x  1500  (BAUMGARTEN). 

Sections  of  pneumonic  lung  should  be  stained  by — 

(a)  Method  of  Gram. 

(b)  Method  of  Friedlander.     This  method  is  employed  to  demonstrate 
the  capsules  in  tissue  sections.     It  consists  in  placing  the  sections  twenty- 
four  hours  in  the  following  solution  : — 

Fuchsine 1 

Distilled  water 100 

Alcohol    ........  5 

Glacial  acetic  acid 2 

They  are  then  rinsed  with  alcohol,  transferred  for  a  couple  of  minutes 
to  a  2  per  cent,  solution  of  acetic  acid,  and  treated  with  alcohol  and  oil 
of  cloves  in  the  usual  way,  and  preserved  in  Canada  balsam. 

Sternberg's  nricrococcus  was  first  found  in  the  blood  of  rabbits 
inoculated  with  saliva.  Three  months  afterwards,  Pasteur  encoun- 
tered the  same  organism  in  rabbits  inoculated  with  the  blood  of  a 
child  suffering  from  rabies.  The  same  organism  in  1883  was  found 
by  Talamon  in  pneumonic  sputum.  It  was  identified  by  Sternberg. 
Two  years  afterwards  further  observations  were  made  by  Frankel, 
Gamaleia,  and  others.  It  has  also  been  found  in  purulent  meningitis 
by  Netter,  and  by  Monti  in  cerebro-spinal  meningitis,  by  Weichsel- 
baum  in  ulcerative  endocarditis,  and  by  others  in  acute  abscess  of 
the  middle  ear,  and  in  purulent  inflammation  of  the  joints  following 
pneumonia. 


236  INFECTIVE    DISEASES. 

Sternberg's  Microeoccus.  (Microbe  de  salive,  Pasteur ;  Mwro- 
coccus  Pasteuri,  Sternberg ;  Lancet-shaped  micrococcus,  Talamoii ; 
Streptococcus  lanceolatus  Pasteuri,  Gamaleia ;  Diplococcus  piwu- 
monice,  Weichselbaum  ;  Bacillus  septicus  sputigenus,  Fliigge  ; 
Micrococcus  of  sputum  septiccemia,  Frankel.)  Spherical  or  oval 
cocci,  singly,  in  pairs  or  in  chains,  often  lanceolate  and  capsuled. 
Stain  readily  with  the  aniline  colours  and  by  Gram's  method  ; 
non-motile.  They  nourish  in  alkaline  media  in  the  incubator.  In 
broth  they  produce  in  twelve  hours  a  cloudiness  due  to  the  develop- 


FIG.  116.— MICROCOCCUS  OP  SPUTUM  SEPTICAEMIA.     From  the  blood  of  a 
Rabbit,     x   1000  (FRANKEL  AND  PFEIFFER). 

ment  of  cocci  and  short  chains.  After  a  time  these  subside  to 
the  bottom  of  the  tube,  and  the  liquid  above  becomes  clear.  In 
plate -cultivations  the  colonies  are  small,  circular,  white,  and  granular. 
In  the  depth  of  gelatine,  minute  white  colonies  develop  along  the 
track  of  the  needle  without  liquefaction  of  the  gelatine ;  and  on 
the  sloping  surface  of  nutrient  agar  or  blood  serum  minute  trans- 
parent drops  appear  along  the  line  of  inoculation.  They  grow  in 
milk,  coagulating  casein ;  but  they  do  not  grow  on  potato.  Sub- 
cultures quickly  lose  their  virulence,  but  regain  it  by  inoculation. 


PNEUMONIA.  237 

The  injection  of  a  minute  quantity  ('2  cc.)  of  a  virulent  culture 
subcutaneously  proves  fatal  to  mice  and  rabbits  in  from  twenty- 
four  to  forty-eight  hours.  Immediately  afterwards  there  is  a  rise  of 
temperature  of  2°  or  3°  0.,  later  it  falls,  and  just  before  death 
it  is  several  degrees  below  normal.  After  death,  the  post-mortem 
appearances  of  septicaemia  are  observed,  in  addition  to  diffuse 
inflammatory  cedema  extending  in  all  directions  from  the  point  of 
injection.  The  subcutaneous  connective  tissue  contains  sanguineous 
serum  and  micrococci  in  abundance.  The  liver  and  spleen  are  some- 


117. — COLONIES  OP  STERNBERG'S  MICROCOCCUS.     Agar  plate-cultivation, 
after  24  hours,     x    100  (FRANKEL  AND  PFEIFFER). 


times  dark  and  engorged,  and  blood  from  the  heart  and  internal 
organs  teems  with  micrococci. 

There  is  no  indication  of  pneumonia  after  subcutaneous  inocula- 
tion, but  intra- pulmonary  injections  produce  fibrinous  pneumonia, 
often  fatal  (Talamon,  Gamaleia).  The  result  is  usually  fatal  in 
rabbits  and  sheep,  but  dogs,  as  a  rule,  recover.  Injection  of  cultures 
into  the  trachea  of  rabbits  is  said  to  induce  typical  pneumonia 
(Monti). 

Sternberg  concludes  that  this  micrococcus  is  the  cause  of  acute 
infectious  pneumonia,  but  the  micrococcus  is  undoubtedly  associated 
with  widely  different  pathological  processes,  and  the  possibility  of  its 
being  a  saprophyte,  which  finds  in  pneumonia  a  suitable  soil  for  its 
development,  must  not  be  overlooked. 


238  INFECTIVE    DISEASES. 

Klein's  Micrococcus. — Klein  found  in  pneumonic  sputum  a 
diplococcus  which  does  not  appear  to  differ  from  Sternberg's  micro  - 
coccus.  In  cover-glass  preparations  the  bacilli  are  surrounded  with 
a  halo,  but  no  definite  capsule,  as  in  Friedlander's  coccus.  They 
appear  as  short  rods  constricted  in  the  centre,  or  dumb-bell  forms, 
and  forms  intermediate  between  cocci  and  bacilli.  In  gelatine,  after 
two  or  three  days,  greyish- white  spots  appear,  which  enlarge  in  the 
next  two  or  three  days  into  flat,  translucent,  greyish-white  plaques, 
with  irregular  serrated  outline.  Colonies  beneath  the  surface  are 
spherical,  and  of  a  brownish-yellow  colour.  In  test-tubes  in  the 
depth  of  the  gelatine  a  whitish-brown  filament  develops  on  incuba- 
tion, composed  of  minute  spherical  colonies,  and  on  the  surface  the 
growth  spreads  out  into  a  greyish-white  film  with  serrated  margin. 
On  the  surface  of  obliquely  solidified  gelatine  the  growth  forms  a 
thin  whitish  film,  which  enlarges  in  breadth  with  irregular  outline, 
reaching  its  maximum  in  about  a  fortnight.  The  growth  on  agar 
is  very  similar.  Broth  becomes  uniformly  turbid  in  twenty-four 
hours,  then  a  powdery  precipitate  makes  its  appearance.  On  potato 
there  is  a  thin,  moist,  faintly  yellowish- brown  film.  Cultures  examined 
in  the  fresh  state  show  many  rods  in  a  resting  stage,  and  others 
actively  motile.  In  addition  to  the  dumb-bell  forms  there  are  others 
of  greater  length,  and  in  old  cultures  involuted  and  degenerated 
forms.  Spore- formation  has  not  been  observed.  A  broth-culture 
inoculated  into  two  rabbits  produced  a  local  tumour  which  subsided 
in  a  week.  Death  ensued  in  one  case  in  eight  days,  and  in  the 
other  in  three  weeks.  There  was  purulent  matter  at  the  seat  of 
inoculation  in  one  ;  in  the  other,  pericardial  exudation  and  hyperaemia 
of  the  lungs.  Broth- cultures  inoculated  intravenously  produced  no 
effect.  In  guinea-pigs  there  was  swelling  at  the  seat  of  inoculation, 
or  slight  indication  of  disease  and  recovery.  Cultures  inoculated 
in  mice  produced  rapid  breathing,  drowsiness,  and  death  in  from 
twenty-four  to  ninety-six  hours.  The  internal  organs  were  con- 
gested, the  lungs  inflamed,  and  the  blood  and  organs  in  the  inoculated 
animals  contained  the  diplococci  in  considerable  numbers. 

Foa  isolated  a  coccus  which  he  named  the  Micrococcus  lanceolatus 
capsulatus.  It  produced  in  small  animals  either  rapid  septicaemia 
arid  death,  or  local  oedema  and  death  at  a  later  period. 

Protective  Inoculation.— Immunity  has  been  produced  in 
rabbits  by  the  intravenous  injection  of  the  virus  in  a  diluted  form. 
Blood  obtained  from  immunised  rabbits  was  kept  at  10°  C.  for  twelve 
hours,  and  then  filtered,  and  animals  injected  with  it  acquired 
immunity  against  virulent  cultures  (Emmerich). 


INFECTIOUS   PLEURO-PNEUMONIA. 


239 


Filtered  cultures  are  said  to  confer  immunity  for  six  months,  and 
raising  the  temperature  of  filtered  cultures  increases  the  strength  of 
the  substance  which  gives  immunity  (Klemperer).  The  blood  serum 
of  immune  animals  can  confer  immunity  on  other  animals,  and,  it  is 
said,  will  arrest  the  progress  of  the  disease  produced  by  injection 
of  healthy  animals  with  virulent  cultures.  The  cultures  contain  a 
proteid  body,  for  which  the  name  pneumo-toxin  has  been  suggested, 
and  anti- pneumo-toxin  has  been  isolated  from  immunised  blood 
serum. 

INFECTIOUS  PLEURO-PNEUMONIA. 

Infectious  pleuro-pneumonia  is  a  highly  infectious  disease 
peculiar  to  cattle ;  it  is  characterised  by  rise  of  temperature  and 
exudation  into  the  lungs.  It  is  often  fatal,  and  sometimes  exists  in 
an  extremely  chronic  form.  It  is  believed  to  have  been  unknown  in 
England  previously  to  1840,  and  is  supposed  to  have  been  introduced 
from  Holland,  where  in  one  year  it  destroyed  seven  thousand  cattle. 


FIG.  118.— ACUTE  CATARRHAL  PNEUMONIA  (Ox). 

<(,  Coagulated  mucus  with  catarrhal  cells  (c)  embedded  in  it ;  b,  catarrhal  cells 
sprouting  from  alveolar  wall,     x  480.     (Hamilton.) 

The  disease  cannot  be  conveyed  artificially.  A  living,  diseased 
animal  must  be  the  medium  of  infection.  The  disease  is  apparently 
only  communicated  by  cohabitation.  Brown  injected  large  quanti- 
ties of  lymph  from  diseased  lungs  into  the  jugular  vein,  into  the 


240  INFECTIVE    DISEASES. 

tissue  of  the  lungs,  and  into  the  trachea,  without  any  result  except 
a  small  abscess  at  the  seat  of  puncture.  Administration  of  the 
virus  by  the  mouth  gave  equally  negative  results.  The  kings  from 
a  recently  killed  animal  infected  with  pleuro-pneumonia  were  placed 
in  a  shed  occupied  by  healthy  heifers,  and  left  there  for  several  days. 
Fodder,  litter,  and  manure  were  taken  from  places  in  which  there 
were  diseased  cattle,  and  placed  in  contact  with  healthy  cattle,  and  sub- 
sequently all  the  animals  used  in  these  experiments  were  slaughtered 
and  carefully  examined,  and  the  results  were  absolutely  negative. 
Similarly  negative  results  followed  experiments  made  by  Sander- 


FIG.  119.— INFECTIOUS  PLEURO-PNEUMONIA  OF  CATTLE,    x   480. 

a, a, a,  Exudation  in  air-vesicles,  composed  of  a  network  of  fibrinous  lymph  with 
entangled  leucocytes;  b,b,  the  same  caseating ;  c,  the  air- vesicle  filled  with 
leucocytes  only.  In  the  centre  is  a  blood-vessel  filled  with  a  fibrinous  plug. 
(Hamilton. ) 

son  and  Duguid,  and  thus  confirmed  the  conclusion  arrived  at  by 
Brown,  that  the  disease  could  only  be  communicated  by  actual  contact 
of  a  living,  diseased  animal  with  a  healthy  one. 

The  symptoms  of  the  disease  in  cattle  are  a  rise  of  temperature 
to  105°  or  107°,  and  a  peculiar  dry  cough,  and  later  the  usual 
indications  of  pneumonia,  difficulty  in  breathing,  and  dulness  on 
percussion.  As  a  rule,  death  follows  from  exhaustion ;  but  the  disease 
may  also  assume  a  chronic  form,  if  the  animal  escapes  slaughter, 
and  the  lung  may  become  gangrenous  or  tubercular.  The  period  of 
incubation  is  about  thirty  days,  but  it  is  uncertain.  The  lesions  are 


FIG.  120.— INFECTIOUS  PLEURO-PNEUMONIA  OF  CATTLE,  x  50. 
a,  a,  a,  Spaces  in  deep  layer  of  pleura  and  interlobular  septa  filled  with  fibrinou^ 
lymph  ;  6,  deep  layer  of  pleura  running  down  to  an  interlobular  septum ;  c,c, 
air-vesicles  filled  with  fibrinous  lymph ;  d,  blood-vessels  of  alveolar  walls, 
much  congested  ;  e,  large  congested  blood-vessels ;  /,/,  interlobular  septa 
infiltrated  with  fibrinous  lymph  ;i  g,  blood-vessel  in  interlobular  septum 
(Logwood,  Eosin  and  Farrant's  solution). — HAMILTON. 

16 


242  INFECTIVE    DISEASES. 

limited  almost  entirely  to  the  lungs  ;  congestion  is  quickly  followed 
by  inflammation  and  effusion  into  the  air  vesicles  and  the  intra- 
lobular  fibrous  tissue  which  is  so  well  marked  in  the  lungs  of  cattle. 
Leucocytes  are  entangled  in  the  fibrinous  lymph,  and  the  intra- 
lobular  septa  are  enormously  enlarged,  so  that  the  red  lobules  are 
mapped  out  by  the  paler  septa,  and  produce  on  section  of  the  diseased 
parts  a  very  striking  marbled  appearance.  A  somewhat  similar 
appearance  is  sometimes  observed  in  septic  pleuro-pneumonia  in 
calves.  The  effusion  occurs  also  in  the  air  vesicles.  The  stages 
of  grey  hepatisation  and  suppurative  softening  have  not,  as  a  rule, 
time  to  develop.  Hsemorrhagic  infarctions  are  sometimes  produced, 
which  in  turn  become  gangrenous  or  cheesy,  and  a  capsule  may  form 
round  the  diseased  part.  Roy  found  micro-organisms  in  the  lymph, 
but  attached  no  importance  to  them.  Bruylants  and  Yerriet  also 
•described  a  micro-organism  in  the  lymph.  Later,  Poels  and  Nolen 
isolated  a  micrococcus  resembling  JFriedlander's  pneumococcus. 
Inoculation  in  the  lungs  produced  a  condition  in  cattle  which  they 
considered  indicative  of  pleuro-pneumonia. 

Lustig  was  unable  to  confirm  these  observations,  but  succeeded 
in  isolating  from  lymph  a  bacillus  and  three  species  of  micrococci. 
•One  of  the  micrococci  formed  an  orange  growth  when  cultivated, 
and  was  regarded  as  the  specific  micro-organism,  as  it  caused  sub- 
cutaneous tumefaction,  and,  it  is  said,  some  degree  of  immunity. 

Brown  cultivated  a  number  of  organisms  which  on  inoculation 
only  produced  local  irritation.  Intravenous  injection  produced  death 
from  septicaemia  in  one  case  in  thirty- six  hours. 

Arloing  isolated  four  different  organisms,  including  a  bacillus 
which  was  named  Pneumo-bacillus  liquefaciens  bovis.  Later,  he 
prepared  a  fluid  from  broth- cultures,  pneumo-bacillin,  which  pro- 
duced a  more  marked  rise  in  temperature  in  animals  suffering  from 
pleuro-pneumonia  than  in  healthy  animals,  and  its  use  was 
suggested  as  an  aid  in  diagnosis.  Arloing  named  the  micro- 
organisms provisionally  Pneumo-bacillus  liquefaciens  bovis,  Pneumo- 
<coccus  gutta  cerei,  Pneumococcus  lichenoides,  and  Pneumococcus 
flavescens. 

.  Pneumo-bacillus  liquefaciens  bovis. — Short  rods,  non- 
motile  ;  spore-formation  not  observed.  They  rapidly  liquefy  gelatine, 
and  form  on  potato  a  white  layer,  which  becomes  brownish  and 
sometimes  greenish.  According  to  Arloing  pure-cultures  produce 
in  the  ox,  when  injected  subcutaneously  or  in  the  lung,  the  same 
lesions  which  are  produced  by  virulent  lymph.  Guinea-pigs  and 
rabbits  are  slightly  susceptible,  dogs  are  immune. 


INFECTIOUS   PLEURO-PNEUMONIA.  243 

Nocard  does  not  accept  Arloing's  conclusions,  and  expresses  the 
opinion  that  the  virus  is  participate,  but  is  not  due  to  any  micro- 
organism which  can  be  detected  or  cultivated  by  the  methods 
at  present  adopted.  In  the  opinion  of  the  author,  who  has 
also  examined  the  micro-organisms  in  pleuro-pneumonia,  it  is 
fully  justifiable  to  regard  the  nature  of  the  contagium  as 
unknown. 

Preventive  Inoculation. — In  1852  Willems  introduced  inocula- 
tion. The  liquid  from  the  lungs  of  an  animal  with  pleuro-pneumonia, 
which  had  recently  died,  was  inoculated  in  the  extremity  of  the  tail 
by  a  puncture  with  a  lancet.  Swelling  occurred  at  the  seat  of  inocu- 
lation, and  on  recovery  the  animals  were  believed  to  be  protected. 
A  Dutch  Commission  reported  that  the  inoculation  gave  a  temporary 
protection.  A  Belgian  Commission  in  the  following  year  reported 
that  the  phenomena  of  inoculation  could  be  produced  several  times 
in  succession  in  the  same  animal,  and  that  it  was  not  a  certain 
preventive.  A  French  Commission  in  1854  concluded  that  a  power 
of  resisting  infection  was  given,  but  the  period  was  undetermined. 
Protective  inoculation  continued  to  be  employed,  and  various  modi- 
fications of  the  method  were  introduced.  Threads  soaked  in  lymph 
were  inoculated,  or  the  lymph  subcutaneously  or  intravenously 
injected. 

The  usual  result  of  the  inoculation  is  swelling  and,  in  about  ten 
or  fourteen  days,  effusion  of  straw-coloured  fluid,  which  is  occasion- 
ally blood-stained.  Gangrene  may  follow,  involving  amputation  of 
the  tail.  Germont  and  Loire  in  Queensland  adopted  the  method — 
which  was  suggested  by  Pasteur — of  inoculating  calves  in  the  loose 
cellular  tissue  behind  the  shoulder.  This  produces  intense  oedema 
and  a  quantity  of  lymph.  There  has  been  much  controversy  with 
regard  to  the  value  of  protective  inoculation. 

Stamping-out  System. — Brown  maintains  that  pleuro- 
pneumonia  can  be  exterminated  only  by  slaughter  of  the  diseased 
animals,  and  quotes  the  results  experienced  in  the  Netherlands  in 
support  of  his  views. 

In  1871  slaughter  for  pleuro-pneumonia  was  commenced  in  the 
Netherlands.  There  were  6,000  cattle  attacked  by  the  disease.  In 
1872  owners  were  compelled  to  slaughter  not  only  diseased  cattle, 
but  those  which  had  been  in  contact  with  them,  unless  inoculated, 
and  the  attacks  were,  in  consequence,  reduced  to  4,000.  In  1873  it 
was  forbidden  to  move  cattle  out  of  infected  districts,  and  the  attacks 
were  reduced  to  2,479.  In  1876  slaughter  of  the  whole  herd  was 
decreed,  and  during  the  first  year  of  this  heroic  system  the  cases  fell 


244 


INFECTIVE   DISEASES. 


from  2,227  in  1875,  to  1,723  in  1876,  to  951  in  1877,  to  698  in 
1878,  to  157  in  1879,  and  to  48  in  1880. 

In  England  the  Pleuro-pneumonia  Act  came  into  force  on 
September  1st,  1890.  Notification  was  to  be  given  by  the  owner 
to  a  police  constable  of  the  district,  who  was  required  to  transmit 
the  information  to  the  Local  Authority  and  also  to  the  Board  of 
Agriculture.  An  inspector,  with  the  aid  of  the  veterinary  surgeon, 
arranged  for  the  slaughter  of  the  suspected  animal,  and,  if  the 
disease  proved  to  be  pleuro- pneumonia,  of  the  rest  of  the  herd.  The 
results  are  shown  in  the  following  table  : — 


YEARS. 

Number 
of 
Infected 
Counties. 

Number 
of 
Fresh 
Outbreaks. 

Number 
of 
Cattle 
Attacked. 

Diseased  Cattle. 

Healthy 
Cattle  in 
contact 
slaughtered. 

Cattle 
slaughtered 
as  suspected, 
but  found 
free  from 
Pleuro- 
Pneumonia. 

Killed. 

Died. 

1890 

36 

465 

2,057 

2,022 

37 

11,301 

1891 

27 

192              778 

778 

— 

9,491 

232 

1892 

10 

35 

134 

134 

— 

3,477 

188 

1893 

4 

9 

30 

30 

— 

1,157 

86 

1894 

2 

2 

15 

15 

— 

391 

41 

Thus  the  number  of  cases  was  reduced  from  2,057  in  1890  to 
15  in  1894. 

A  departmental  committee  appointed  in  1892  to  inquire  into 
pleuro-pneumonia  and  tuberculosis,  came  to  the  following  conclusions 
with  regard  to  pleuro-pneumonia  :— 

(1)  That  the  system  of  compulsory  slaughter  be  applied  not  only  to 
all  diseased  cattle,  but  also  to  all  cattle  which  have  been  in  association 
with  them,  or  otherwise  in  any  manner  exposed  to  the  infection  of  the 
disease. 

(2)  Compulsory  slaughter  should  be  accompanied  by  supplementary 
measures,  such  as  restrictions  on  the  movement  and  sale  of  cattle  within. 
or  coming  from,  infected  districts. 

(3)  Any  exception  to,  or  modification  of,  the  system  of  compulsory 
slaughter,   as   provided  in   the    Slaughter  Order,    1888,   should    only    be 
applicable  to  cattle  in  the  dairy   yards,  byres,    and   cowsheds   of   large 
towns,  the  owners  or  occupiers  of  which  may  claim  in  writing  the  privi- 
lege of  exemption  for  their  cattle   from   immediate   slaughter,   on   the 
following  conditions  : — 


INFECTIOUS   PLEURO-PNEUMONIA.  245 

(a)  No  head  of  cattle  that  has  been  brought  into  such  dairy  premises 
shall  be  removed  therefrom,  except  for  the  purpose  of  immediate 
slaughter. 

(6)  In  the  event  of  an  outbreak  of  pleuro-pneumonia,  all  the  diseased 
cattle  shall  be  slaughtered. 

(c)  All  the  remaining  cattle  on  such  premises  where  an  outbreak  has 
occurred  shall  be  branded,  and  regularly  subjected  to  the  ther- 
mometer test ;  and  whenever  a  continuous  increase  of  temperature, 
rising  above  104°,  is  shown,  they  shall  be  slaughtered. 

(r/)  No  fresh  cattle  shall  be  admitted  into  such  premises  while  any 
of  the  cattle  thus  branded  remain  alive. 

(4)  Inoculation  cannot  be  recommended  as  a  means  of  eradicating 
pleuro-pneumonia,  nor  as  practicable  under  existing  conditions.  Although 
it  is  open  to  owners  to  inoculate  their  cattle,  it  should  be  distinctly 
understood  that  that  operation  shall  not  give  them  any  immunity  from 
the  regulations  above  suggested. 

The  order  at  present  in  force  is  the  Pleuro-Pneumonia  Order 
of  1895.  In  addition  to  regulations  for  the  movement  of  cattle, 
for  disposal  of  carcasses,  for  markets,  and  for  compensation  for 
slaughter,  the  Order  contains  the  following  provisions  : — 


NOTIFICATION. 
Notice   of  Disease. 

(1)  Every  person  having  or  having  had  in  his  possession  or  under  his 
charge  a  head  of  cattle  affected  with  or  suspected  of  pleuro-pneumonia 
shall  with  all  practicable  speed  give  notice  of  the  fact  of  the  head  of  cattle 
being  so  affected  or  suspected  to  a  constable  of  the  police  force  for  the 
police  area  wherein  the  head  of  cattle  so  affected  or  suspected  is  or  was. 

(2)  The  constable  receiving  such  notice  shall  immediately  transmit 
the  information  by  telegraph  to  the  Board  of  Agriculture. 

(3)  The  constable  shall  also  forthwith  give  information  of  the  receipt 
by  him  of  the  notice  to  an  Inspector  of  the  Local  Authority,  who  shall 
forthwith  report  the  same  to  the  Local  Authority. 

Duty  of  Inspector  to  act  immediately. 

(1)  An  Inspector  of  a  Local  Authority  on  receiving  in  any  manner 
whatsoever  information  of  the  supposed  existence  of  pleuro-pneumonia, 
or  having  reasonable  ground  to  suspect  the  existence  of  pleuro-pueumonia, 
shall  proceed  with  all  practicable  speed  to  the  place  where  such  disease, 
according  to  the  information  received  by  him,  exists,  or  is  suspected  to 
exist,  and  shall  there  and  elsewhere  put  in  force  and  discharge  the  powers 
and  duties  conferred  and  imposed  on  him  as  Inspector  by  or  under  the 
Act  of  1894  and  this  Order. 

(2)  The  Inspector  shall  forthwith  report  to  the  Board  of  Agriculture. 


246  INFECTIVE   DISEASES. 

No  Movement  into  or  out  of  Pleura-pneumonia  Infected  Place  without 

Licence. 

Cattle  shall  not  be  moved  into  or  out  of  an  Infected  Place  except  with 
a  Movement  Licence  of  an  Inspector  or  officer  of  the  Board,  and  such 
cattle  shall  not  be  moved  except  in  accordance  with  the  conditions 
contained  in  such  Licence. 

Pleuro-pneumonia  found  in  a  Market,  Railway  Station,  Grazing  Park, 
or  other  like  Place,  or  during  Transit. 

The  Inspector  of  the  Local  Authority  shall  cause  to  be  seized  all  the 
cattle  affected  with  pleuro-pneumonia,  and  also  all  cattle  being  in  or  on 
the  market,  fair,  sale-yard,  place  of  exhibition,  lair,  landing-place,  wharf, 
railway  station,  common,  uninclosed  land,  farm,  field,  yard,  shed,  park,  or 
other  such  place  as  aforesaid,  and  shall  forthwith  transmit  the  information 
by  telegraph  to  the  Board  of  Agriculture. 

The  Inspector  of  the  Local  Authority  shall  cause  all  such  cattle  so 
seized  to  be  detained  at  the  place  where  they  are  seized,  or  to  be  moved 
to  some  convenient  and  isolated  place  and  there  detained. 

Removal  of  Dung  or  other  Things. 

It  shall  not  be  lawful  for  any  person  to  send  or  carry,  or  cause  to  be 
sent  or  carried,  on  a  railway,  canal,  river,  or  inland  navigation,  or  in  a 
coasting  vessel,  or  on  a  highway  or  thoroughfare,  any  dung,  fodder,  or 
litter  that  has  been  in  an  Infected  Place,  or  that  has  been  in  any  place  in 
contact  with  or  used  about  a  diseased  or  suspected  head  of  cattle,  except 
with  a  Licence  of  an  Inspector  or  officer  of  the  Board  or  of  an  Inspector 
of  the  Local  Authority. 

Report  to  Board  of  Cattle  that  have  been  in   Contact  with  Cattle   affected 
with  Pleuro-pneumonia. 

Where  it  appears  to  a  Local  Authority  that  there  is  within  their  District 
any  head  of  cattle  which  has  been  in  the  same  field,  shed,  or  other  place, 
or  in  the  same  herd,  or  otherwise  in  contact,  with  any  head  of  cattle 
affected  with  pleuro-pneumonia,  or  otherwise  exposed  to  the  infection 
thereof,  the  Local  Authority  shall  forthwith  report  the  facts  of  the  case 
to  the  Board  of  Agriculture. 

Disinfection. 

An  Inspector  or  officer  of  the  Board  may  cause  or  require  any  shed  or 
other  place  which  has  been  used  for  a  head  of  cattle  while  affected  with 
or  suspected  of  pleuro-pneumonia,  and  any  utensil,  pen,  hurdle,  or  other 
thing  used  for  or  about  such  head  of  cattle,  to  be  cleansed  and  disinfected 
to  his  satisfaction. 

Occupiers  to  give  Facilities  for  Cleansing. 

(1)  The  owner  and  occupier  and  person  in  charge  of  any  shed  or  other 
place  which  has  been  used  for  any  head  of  cattle  while  affected  with  or 
suspected  of  pleuro-pneumonia  shall  give  all  reasonable  facilities  to  an 


INFLUENZA.  247 

Inspector  or  officer  of  the  Board  for  the  cleansing  and  disinfection  of  such 
place,  and  of  any  utensils,  pens,  hurdles,  or  other  things  used  for  or  about 
such  cattle. 

(2)  Any  person  failing  to  comply  with  the  provisions  of  this  Article 
shall  be  deemed  guilty  of  an  offence  against  the  Act  of  1894. 

Prohibition  to  Expose  or  Move  Diseased  or  Suspected  Cattle. 

(1)  It  shall  not  be  lawful  for  any  person — 

(a)  To  expose  a  diseased  or  suspected  head  of  cattle  in  a  market  or 
fair,  or  in  a  sale-yard,  or  other  public  or  private  place  where  cattle 
are  commonly  exposed  for  sale  ;  or 

(6)  To  place  a  diseased  or  suspected  head  of  cattle  in  a  lair  or  other 
place  adjacent  to  or  connected  with  a  market  or  a  fair,  or  where 
cattle  are  commonly  placed  before  exposure  for  sale  ;  or 

(?)  To  send  or  carry,  or  cause  to  be  sent  or  carried,  a  diseased  or 
suspected  head  of  cattle  on  a  railway,  canal,  river,  or  inland 
navigation,  or  in  a  coasting  vessel  ;  or 

(f/)  To  carry,  lead,  or  drive,  or  cause  to  be  carried,  led,  or  drivenr 
a  diseased  or  suspected  head  of  cattle  on  a  highway  or  thorough- 
fare :  or 

(e)  To  place  or  keep  a  diseased  or  suspected  head  of  cattle  on  common 
or  uninclosed  land,  or  in  a  field  or  place  insufficiently  fenced,  or 
in  a  field  adjoining  a  highway  unless  that  field  is  so  fenced  or 
situate  that  cattle  therein  cannot  in  any  manner  come  in  contact 
with  cattle  passing  along  that  highway  or  grazing  on  the  sides 
thereof  ;  or 

(/)  To  graze  a  diseased  or  suspected  head  of  cattle  on  pasture  being 
on  the  sides  of  a  highway  ;  or 

((/)  To  allow  a  diseased  or  suspected  head  of  cattle  to  stray  on  a 
highway  or  thoroughfare  or  on  the  sides  thereof  or  on  common 
or  uninclosed  land,  or  in  a  field  or  place  insufficiently  fenced. 

INFLUENZA. 

Influenza  is  an  infectious  disease  characterised  by  a  catarrh  of 
the  respiratory  or  the  gastric  mucous  membrane,  accompanied  by 
great  prostration  and  mental  depression,  and  frequently  ending 
fatally  by  pneumonic  complication.  One  attack  is  not  protective. 
The  disease  has  occurred  in  the  form  of  great  epidemics,  like  the 
pandemic  of  1890,  which  is  said  to  have  started  from  Bokhara,  and 
travelled  to  St.  Petersburg,  Berlin,  Paris,  and  London,  whence  it 
spread  all  over  this  country.  The  incubation  period  is  extremely 
short,  only  a  few  hours,  so  that  numbers  are  attacked  almost 
simultaneously.  The  occurrence  of  cases  in  succession  in  a  family, 
the  importation  of  the  disease  by  an  infected  person,  and  the  escape 
of  persons  in  completely  isolated  localities,  point  to  the  existence 
of  a  living  contagium.  Pfeiffer  claims  to  have  identified  it  with  a 


248  INFECTIVE    DISEASES. 

bacillus  which  was  found  by  him  in  the  purulent  bronchial  secretion, 
and,  by  Canon,  in  the  blood. 

Bacillus  of  Influenza. — Very  small  rods,  singly  or  in  leptothrix 
filaments.  They  stain  with  the  aniline  dyes,  but  not  by  Gram's 
method.  They  are  non- motile  and  aerobic ;  they  do  not  grow  in 
gelatine  at  the  temperature  of  the  room.  On  glycerine-agar  very 
small  transparent  drop-like  colonies  develop  in  about  twenty-four 
hours.  In  broth  there  is  only  a  very  scanty  growth  of  whitish 
particles  on  the  surface,  which  subside  and  form  a  woolly  deposit. 
They  are  found  especially  in  the  bronchial  secretion,  and  only  in 
cases  of  influenza.  Canon  obtained  them  by  puncturing  the  finger, 


FIG.  121. — BACILLUS  OF  INFLUENZA. 
From  a  culture  on  gelatine,  x  1000.     (!TZEROTT  AND  NIEMANN.  ) 

and  allowing  a  few  drops  of  the  blood  to  fall  upon  the  surface  of 
glycerine-agar  in  a  Petri's  dish.  The  organism  will  retain  its 
vitality  for  fourteen  days  in  sputum,  but  is  quickly  detroyed  by 
drying.  It  is  said  that  by  applying  the  bacillus  to  the  nasal 
mucous  membrane  in  monkeys,  symptoms  similar  to  influenza  were 
produced. 

METHOD  OF  STAINING. 

To  stain  the  bacilli  use  Neelseri's  solution  or  Loftier' s  methylene- 
blue ;  or  the  following  method  : — 

Canon's  method. — Spread  the  blood  on  cover- glasses,  allow  them 
to  dry,  immerse  for  five  minutes  in  absolute  alcohol,  and  stain  in  the 
following  solution : — 


INFLUENZA.  249 

Aqueous  solution  of  methylene-blue,  strong,  40  parts ;  |  per 
cent,  solution  of  eosin  in  70  per  cent,  alcohol,  20  parts ;  distilled 
water,  40  parts. 

Float  the  cover-glasses  from  three  to  six  hours  in  a  capsule 
placed  in  the  incubator  at  37°  C.,  wash  with  water,  and  dry  and 
mount  in  balsam.  The  red  corpuscles  will  be  stained  pink,  and 
the  leucocytes,  with  the  bacilli  in  them,  blue. 


FIG.  122.— BACILLUS  OF  INFLUENZA. 

From  a  cultivation  showing  filaments  composed  of  long  and  short  rods, 
cocci-forms  and  irregular  elements,     x  1200. 

EQUINE  INFLUENZA. 

Equine  influenza,  or  "  pink-eye,"  has  been  noficed  to  be  prevalent 
at  the  same  time  as  epidemics  of  influenza  in  man,  but  there  does 
not  appear  to  be  any  evidence  of  intercommunicability  or  of  any 
relation  between  the  two  diseases. 


CHAPTER   XVIII. 

ORIENTAL  PLAGUE. — RELAPSING   FEVER. — TYPHUS  FEVER. — YELLOW 

FEVER. 

THE  PLAGUE. 

THE  plague  is  a  highly  infectious  disease,  having  its  origin  in 
putrefaction  and  filth,  in  tropical  climates.  The  virus  in  its  effects 
resembles  that  of  typhus.  The  period  of  incubation  varies  from  a 
few  hours  to  a  week.  The  disease  produces  high  temperature  and 
decomposition  of  the  blood,  and  dark  hsemorrhagic  patches  appear 
on  the  skin,  but  there  is  no  eruption.  Lymphatic  inflammation  and 
buboes  almost  invariably  occur.  The  virus  is  intensified  by  warmth 
and  overcrowding  in  houses,  and  dissipated  by  exposure  to  fresh  air. 

When  the  plague  occurred  in  this  country  it  was  recognised  as  a 
foreign  pestilence  from  the  East,  and  once  imported  it  was  fostered 
and  intensified  in  virulence  wherever  there  was  filth,  putrefaction, 
and  overcrowding.  The  disease,  like  the  small-pox,  was  communicated 
from  one  person  to  another.  If  a  case  occurred  in  a  house  other 
inmates  were  liable  to  suffer  from  the  disease,  while  visitors  to  the 
house  ran  a  similar  but  less  risk.  There  was  a  good  deal  of  variation 
both  in  the  infectivity  of  the  virus  and  in  the  susceptibility  of 
individuals,  so  that  one  contemporary  writer  remarked  that  "  no 
one  can  account  for  how  it  comes  to  pass  that  some  persons  shall 
receive  the  infection  and  others  not." 

Medical  men  were  credited  with  enjoying  an  extraordinary  degree 
of  immunity,  though  there  were  members  of  the  medical  profession 
who  undoubtedly  died  of  the  plague.  This  tradition  has  been 
supported,  to  a  certain  extent,  by  the  experience  of  the  plague 
in  modern  times.  In  the  epidemic  in  Egypt,  in  1835,  of  the  ten 
French  physicians  engaged  there,  only  one  died  ;  and  while  those 
who  buried  the  victims  of  the  plague  were  liable  to  suffer  from 
it,  and  many  did  so,  yet  the  medical  men  made  more  than  one 
hundred  post-mortem  examinations  without  any  death  resulting. 

250 


THE    PLAGUE.  251 

The  clothes  arid  coverings  of  the  infected  often  spread  the  disease,, 
and  yet  there  are  numerous  examples  of  persons  who  without  having 
adopted  any  method  of  protection  occupied  the  beds  of  plague  patients- 
without  contracting  the  malady. 

The  plague  is  transmissible  from  one  country  to  another 
by  sea.  An  infected  ship  becomes  an  infective  centre  as  readily 
as  an  infected  house.  Once  imported,  whether  by  land  or  sea,  the 
virus  from  infected  persons  or  merchandise  spreads  wherever  the 
environment  is  favourable  for  its  development  and  extension. 

Old  London  afforded  in  every  way  a  suitable  environment  for 
the  plague.  The  situation  of  the  city  was  unhealthy,  and  the  old 
town  ditch  was  a  receptacle  for  all  kinds  of  filth.  The  houses' 
projected  over  the  roadway,  and  the  streets  were  saturated  with 
constant  contributions  of  slops  and  of  excrement  from  animals  and 
human  beings.  The  houses  were  often  filthy  and  unventilated,  and 
the  floors  strewn  with  rushes,  which  were  seldom  changed.  Erasmus 
goes  so  far  as  to  say  that  the  rushes  were  piled  the  new  iipon  the 
old  for  twenty  years,  and  were  fouled  with  spilliiigs  of  beer, 
fragments  of  fish,  expectoration,  vomit,  excrement,  and  urine. 
Another  very  striking  insanitary  feature  of  Old  London  was  the 
overcrowded  state  of  the  graveyards,  which  was  well  calculated  to 
predispose  to  pestilence,  if  not  actually  to  produce  it.  The  burials 
were  so  frequent  in  St.  Paul's  Churchyard  that  a  new  grave  could 
scarcely  be  dug  without  bodies  being  exposed  in  all  stages  of 
putrefaction. 

In  1894  the  plague  broke  out  in  China,  with  all  the  symptoms 
of  the  fatal  bubonic  pest  of  Old  London.  The  disease  was  confined 
to  the  poorest  classes  and  the  most  overcrowded  and  most  filthy 
localities.  In  Canton  the  deaths  exceeded  one  hundred  thousand, 
and  in  Hong-Kong  numbered  about  ten  thousand.  The  disease 
was  contagious,  and  mainly  diffused  by  personal  contact.  Death 
occurred,  as  a  rule,  in  from  twenty-four  hours  to  five  days.  The 
English  and  European  community  escaped,  with  the  exception  of  a 
very  few  out  of  a  large  number,  mostly  soldiers,  employed  in  cleansing 
the  houses.  The  disease  was  a  specific  fever,  intensely  fatal,  accom- 
panied by  high  temperature,  cerebral  congestion,  delirium,  and  the 
formation  of  buboes.  The  buboes  consisted  of  exquisitely  painful  and 
swollen  lymphatic  glands.  All  the  glands,  in  some  cases,  were  affected. 

According  to  Cantlie  the  glandular  swelling  when  first  recognised 
was  almond-shaped  in  the  inguinal  region,  and  globular  in  other 
regions,  with  peri-glandular  oedema.  The  swelling  rapidly  increased 
in  size,  becoming  softer',  less  definite  in  outline,  and  less  tender,  until 


OF  THB 

UNIVERSITY 


252  INFECTIVE    DISEASES. 

by  the  end  of  five  or  six  days  it  consisted  of  an  elevated  mass,  doughy 
to  the  touch,  almost  circular,  with  a  diameter  of  six  inches.  The 
skin  over  the  swelling  was  livid  and  dimpled.  The  swelling  was  in 
some  cases  due  to  purulent  effusion,  but  more  frequently  on  incision 
there  was  only  an  escape  of  sero-sangumeous  fluid.  The  cervical 
glands  in  very  severe  cases  sometimes  attained  an  enormous  size. 

Three  out  of  seven  Japanese  medical  men  were  attacked  and  one 
died,  but  none  out  of  eleven  English  doctors,  though  they  were  equally 
exposed  to  infection.  Of  eight  Englishmen  attacked  seven  were  among 
the  soldiers  employed,  and  only  two  died.  No  nurses  or  attendants 
on  the  sick  were  attacked.  The  virus  appeared  to  be  intimately 
connected  with  filth  in  the  soil.  According  to  the  Chinese,  rats, 
poultry,  goats,  sheep,  cows,  and  buffaloes  are  susceptible.  In  the 
houses  and  hotels  dead  rats  were  found  in  great  numbers  :  it  was  said 
that  they  emerged  from  their  haunts  in  sewers  and  drains,  appeared 
to  be  dazed,  and  limped  about,  owing  to  the  formation  of  buboes  in 
their  hind  legs.  Rats,  mice,  and  guinea-pigs  inoculated  with  virus 
from  a  human  lymphatic  gland  died  with  development  of  buboes. 
It  appears  to  be  clearly  proved  that  rats  suffer  from  the  plague  in 
common  with  man,  and  it  has  also  been  suggested  that  they  may 
serve  to  spread  the  disease.  Bacilli  were  found  in  human  blood  and 
in  the  swollen  lymphatic  glands  by  Kitasato,  and  independently  by 
Yersin. 

Bacillus  of  Plague. — Short  rods  with  rounded  ends.  They 
stain  with  aniline  dyes,  but  not  by  Gram's  method.  The  stain 
collects  at  the  ends  of  the  rods,  leaving  a  clear  space  in  the  middle. 
Sometimes  the  rods  are  surrounded  by  a  capsule.  They  are  found 
in  abundance  in  the  buboes,  and  in  small  numbers  in  the  blood  in 
very  serious  and  rapidly  fatal  cases. 

Material  from  the  buboes  inoculated  on  agar  gives  rise  to  white 
transparent  colonies,  which  have  an  iridescent  edge  when  examined 
by  reflected  light. 

The  bacilli  grow  more  readily  on  glycerine-agar  and  on  solidified 
serum.  In  broth  cultures  the  liquid  remains  clear,  and  a  flocculent 
deposit  forms  on  the  sides  and  at  the  bottom  of  the  vessel. 

An  alkaline  solution  of  peptone  2  per  cent.,  with  from  1  to  2  per 
cent,  of  gelatine,  is  the  best  nutrient  medium.  In  cultures  the 
bacilli  develop  chains  of  short  rods  and  well-marked  involution 
forms.  Swollen  and  degenerated  forms  are  found  most  abundantly 
in  old  cultures,  and  stain  with  difficulty. 

Mice,  rats,  and  guinea-pigs,  inoculated  with  bubonic  tissue,  die  in 
a  few  days,  numerous  bacilli  being  found  in  the  lymphatic  glands. 


THE   PLAGUE.  253 

spleen,  and  blood.  Guinea-pigs  die  in  from  two  to  five  days,  and 
mice  in  one  to  three  days. 

In  guinea-pigs  after  some  hours  there  is  oedema  at  the  seat  of 
inoculation,  and  the  lymphatic  glands  are  swollen.  After  twenty  - 
four  hours  the  animal  refuses  to  eat,  has  a  staring  coat,  and  after 
a  time  suddenly  falls  on  its  side,  and  is  attacked  by  convulsions, 
which  become  more  and  more  frequent  until  death  occurs. 

After  death  the  seat  of  inoculation  is  found  to  be  extensively 
codematous,  and  the  neighbouring  .lymphatic  glands  enlarged  and 
filled  with  bacilli.  The  intestine  is  often  congested,  and  the  liver  is 


ra  tm 

IT    >  » 


FIG.  123.— BACILLI  OF  PLAGUE  AND  PHAGOCYTES,     x   800. 
From  human  lymphatic  gland.     (AOYAMA.) 

congested  and  enlarged.  In  less  acute  cases  an  abscess  of  the 
abdominal  wall  occasionally  results. 

The  bacilli  are  sometimes  found  in  the  pleural  and  peritoneal 
exudation.  The  liver  and  spleen  also  contain  many  bacilli.  Those 
in  the  blood  are  a  little  longer  than  those  in  the  lymphatic  glands. 

Inoculations  can  readily  be  made  from  guinea-pig  to  guinea- 
pig  by  using  the  pulp  of  the  spleen,  or  the  blood.  Cultures  lose  their 
virulence  gradually,  but  the  virus  can  be  intensified  by  successive 
inoculations  in  animals.  The  disease  is  infectious  to  mice  as 
well  as  inoculable.  Pigeons  are  insusceptible.  Rats  and  flies  may 
convey  the  bacilli. 

According  to  Aoyama,  the  bacilli  found  in  the  blood  of  plague 


254  INFECTIVE    DISEASES. 

patients  and  in  the  buboes  are  not  identical.  The  bacilli  in  the 
buboes  are  different  in  form,  and  they  stain  by  Gram's  method. 

There  is  no  doubt  that  the  micro-organism  which  was  found  in 
blood  is  very  similar  to  the  bacillus  of  fowl  cholera,  and  it  is  quite 
possible  that  the  so-called  plague  bacillus  is  really  identical  with  the 
bacillus  of  hsemorrhagic  septicaemia,  and  that  the  real  nature  of  the 
contagium  in  bubonic  plague  is  unknown. 

Protective  Inoculation. — Yersin,  Calmette,  and  Borrell  claim 
not  only  to  have  produced  immunity,  but  to  have  cured  animals 
after  infection.  Cultures  on  agar  heated  to  58°  C.  for  an  hour 
were  attenuated,  and  rabbits  after  intravenous  or  subcutaneous 
inoculation  were  protected  against  virulent  cultures.  The  serum 
of  immunised  rabbits  was  capable  of  protecting  from  subsequent 
virulent  cultures,  and  neutralising  the  effect  of  a  previous  inoculation 
of  a  virulent  culture.  A  horse  was  inoculated  with  cultures  which 
killed  mice  in  two  days,  and  after  six  weeks  a  serum  was  obtained 
which  produced  immunity  in  mice  and  guinea-pigs. 

Stamping-out  System.— It  is  not  until  the  sixteenth  century 
that  we  hear  of  preventive  measures  being  attempted  in  England, 
and  then  they  appear  to  have  been  adopted  only  when  an  outbreak 
threatened  to  be  very  serious. 

Early  in  the  sixteenth  century  all  those  who  had  the  plague  in  their 
houses  were  ordered  to  put  up  wisps,  and  to  carry  white  rods  in  their 
hands. 

In  1543  the  Plague  Order  of  Henry  VIII.  was  issued.  In  place  of 
wisps  the  sign  of  the  cross  was  to  be  made  on  every  infected  house,  and 
to  remain  there  for  forty  days.  Persons  afflicted  with  the  disease  were 
to  refrain,  if  possible,  from  going  out  of  doors,  or  for  forty  days  to  carry 
a  white  rod  in  the  hand.  All  straw  from  the  infected  houses  was  to  be 
carried  into  the  fields  and  burnt.  Churchwardens  were  directed  to  keep 
beggars  out  of  churches  on  holy  days,  and  all  streets  and  lanes  were  to 
be  cleansed. 

In  1547  the  means  of  notification  was  a  blue  cross  with  the  addition 
of  the  inscription  Lord  have  mercy  upon  us.  Later  on  the  colour  of  the 
cross  was  changed  to  red. 

With  the  outburst  of  the  plague  in  1563,  came  an  attempt  to  enforce 
a  terrible  system  of  compulsorily  shutting  up  infected  families.  The 
doors  and  windows  in  such  houses  were  to  be  closed,  and  no  inmates  were 
to  leave  the  premises  and  no  visitors  to  be  allowed  for  forty  days,  No 
better  incubator  on  a  large  scale  could  possibly  have  been  devised  for 
both  breeding  and  intensifying  the  virulence  of  the  plague  bacillus,  or 
whatever  may  be  the  contagium  vivum  of  this  disease. 

This  compulsory  shutting  up  of  the  sick  with  the  healthy  amounted 
to  a  compulsory  infection  of  many  of  the  unfortunate  inmates  who  might 


THE    PLAGUE.  255 

otherwise  have  escaped,  and   very  naturally  the  order  was  frequently 
infringed. 

In  1568  the  Lord  Mayor  of  London  drew  up  instructions  for  the 
Aldermen  for  dealing  with  the  plague.  It  was  enacted  that  constables 
and  officers  should  search  out  infected  houses  and  report  to  the  authorities. 
In  other  words,  that  there  should  be  notification  by  the  police.  All  infected 
houses  were  to  be  shut  up,  and  no  person  to  be  allowed  to  come  out  for 
twenty  days.  All  bedding  and  clothes  used  by  the  victims  were  to  be 
destroyed. 

At  Westminster  these  instructions  were  to  be  enforced  under  a  penalty 
of  seven  days  in  the  stocks,  with  imprisonment  to  follow,  making  in  all 
-a  punishment  of  forty  days. 

In  1581  the  Lord  Mayor  transferred  notification  from  the  constables 
to  searchers.  Two  honest  and  discreet  matrons  in  every  parish  were  to 
search  the  body  of  every  such  person  that  happened  to  die  in  the  parish. 
They  were  ordered  to  make  a  true  report  to  the  clerk  of  the  parish,  and 
the  said  clerk  had  to  report  to  the  wardens  of  the  parish.  For  failing 
to  notify,  the  penalty  was  an  exemplary  term  of  imprisonment.  The 
searchers  were  of  course  likely  to  be  offered  heavy  bribes  by  the  people 
to  suppress  reports,  owing  to  their  anxiety  to  avoid  the  shutting  up  of 
infected  houses. 

The  continued  prevalence  of  the  plague  led  to  the  publication,  in 
1593,  of  a  book  by  Simon  Kellwaye.  One  chapter  "teacheth  what 
orders  magistrates  and  rulers  of  citties  and  towns  should  cause  to  be 
observed,"  which  included  among  other  regulations  that  no  dunghills 
were  to  be  allowed  near  the  city,  and  the  streets  were  to  be  watered 
and  cleansed. 

No  surgeons  or  barbers  who  let  blood  were  to  cast  the  same  into  the 
streets.  All  those  visiting  and  attending  the  sick  to  carry  something  in 
their  hand  to  be  known  from  other  people  ;  and  if  the  infection  were  in 
few  places,  all  the  people  were  to  be  kept  in  their  houses  during  the  time 
of  their  visitation,  and  when  this  was  over,  all  clothes,  bedding,  and 
other  such  things  used  upon  the  sick,  were  to  be  burnt. 

In  1603  Thomas  Lodge  recommended  that  discreet  and  skilful  men 
should  be  appointed  in  every  parish  to  notify  sickness  to  the  authorities, 
and  so  cause  them  to  be  visited  by  expert  physicians,  and  that  such  as 
were  sick  should  be  separated  from  the  whole,  and  that  isolation  hospitals 
should  be  built  outside  the  City  in  separate  and  unfrequented  places. 

In  1665  the  Great  Plague  of  London  occurred,  and  was  attributed  by 
some  to  the  importation  of  an  infected  bale  of  silks  from  the. Levant. 
.  According  to  Hodges  the  disease  stayed  among  the  common  people,  and 
hence  was  called  The  Poor's  Plague.  He  criticised  the  system  of  shutting 
up  infected  houses,  and  strongly  recommended  that  those  who  were 
untouched  in  infected  houses  should  receive  "  accommodation  outside  the 
city."  The  sick  were  to  be  removed  to  convenient  apartments  provided 
on  purpose  for  them.  To  quote  his  own  words,  "  Timely  separation  of 
the  infected  from  the  well  is  absolutely  necessary  to  be  done." 

For  the  purification  of  houses  his  directions  were  to  place  "  a  chafing 


256  INFECTIVE   DISEASES. 

dish  in  the  middle  of  the  room,  where  proper  things  were  burnt  and 
exhaled  all  around."     The  use  of  sulphur  and  quicklime  was  mentioned.* 

Preventive  measures  were  drawn  up  and  published  by  the  Lord  Mayor 
and  Aldermen.  Examiners  in  Health,  watchmen,  and  searchers  w.ere 
appointed.  Surgeons  were  selected  to  assist  the  searchers  in  making  their 
reports,  and  a  fee  of  twelve  pence  was  allowed  for  every  case.  The  disease 
was  immediately  notified  to  the  Examiner  of  Health.  Rules  for  disinfec- 
tion were  made,  and  every  infected  house  was  shut  up,  and  no  one  removed 
except  to  a  pest-house  or  tent.  Orders  were  issued  for  cleaning  and  sweep- 
ing the  streets.  Hackney  coaches  were  not  to  be  used  after  conveying 
patients  to  the  pest-house  until  they  had  been  well  aired.  Regulations 
were  also  made  dealing  with  loose  persons,  assemblies,  and  drinking  taverns. 

The  plague  was  scarcely  over  before  the  whole  city  was  in  flames.  A 
new  city  speedily  rose  upon  the  ashes  of  Old  London.  A  few  sporadic 
cases  of  plague  are  given  in  the  London  Bills  of  Mortality  down  to  1679r 
when  they  finally  ceased.  London  was  sterilised  by  the  great  fire.  '*  Great 
as  this  calamity  was,"  wrote  Thomas  Pennant,  "yet  it  proved  the  provi- 
dential cause  of  putting  a  stop  to  one  of  far  more  tremendous  nature. 
The  plague,  which,  for  a  series  of  ages,  had,  with  very  short  intervals^ 
visited  our  capital  in  its  most  dreadful  forms,  never  appeared  there  again 
after  the  rebuilding  of  the  city  in  a  more  open  and  airy  manner  ;  which 
removed  several  nuisances,  which  if  not  the  origin  of  a  plague,  was 
assuredly  one  great  pabulum,  when  it  had  seized  our  streets." 

In  the  years  1720-22  there  was  a  terrible  outburst  of  plague  in  France. 
It  was  attributed  at  Marseilles  to  importation  by  a  ship  from  Syria.  This 
caused  a  panic  in  England,  and  the  Lords  Justices  considered  it  necessary 
for  the  public  safety  that  measures  should  be  taken  to  defend  the  country 
from  a  fresh  invasion  of  this  disease.  Dr.  Richard  Mead  was  entrusted 
with  drawing  up  the  required  recommendations.  Mead  laid  it  down  as 
an  essential  doctrine  that  the  plague  was  not  native  to  this  country,  and 
therefore  the  first  thing  was  to  prevent  importation,  and  if  such  a  misfor- 
tune occurred,  it  was  to  be  prevented  from  spreading.  How  was  this  to 
be  accomplished  ?  Briefly  stated,  his  system  was  as  follows  :  Lazarettoes 
were  to  be  provided  for  the  reception  of  infected  men  and  merchandise. 
The  healthy  were  to  change  their  clothes  and  to  be  kept  in  quarantine,  and 
the  sick  were  to  be  kept  remote  from  the  healthy  and  their  clothes 
destroyed. 

If,  through  a  miscarriage  in  the  public  care,  by  the  neglect  of  officers 
or  otherwise,  the  disease  was  imported,  then  "  the  civil  magistrates  were 
to  make  it  as  much  for  the  interest  of  the  afflicted  families  to  discover 

*  During  outbreaks  of  the  plague  amulets  were  extremely  popular.  Walnuts 
filled  with  mercury,  pieces  of  cloth  coated  with  arsenic,  and  arsenical  cakes, 
were  very  generally  worn.  The  College  of  Physicians  recommended  issues  on 
the  arms  and  legs.  Dr.  Hodges  wrote,  that  the  more  of  the  ulcers  that  were 
made  the  better,  although  their  largeness  answered  as  well  as  more  in  number. 
If:  two  issues  were  preferred,  it  was  recommended  to  make  one  on  the  left  arm 
and  the  other  on  the  opposite  leg.  A  somewhat  similar  plan  was  adopted  in 
Circassia  by  small-pox  inoculators. 


RELAPSING    FEVER.  257 

their  misfortune,  as  it  was  when  a  house  was  on  fire,  to  call  in  the 
assistance  of  the  neighbourhood."  The  shutting  up  of  infected  houses 
was  condemned  in  the  strongest  terms,  and  a  system  of  notification  and 
isolation  was  proposed  on  the  lines  originally  suggested  by  Dr.  Hodges. 

1 .  .1  Council  of  Health  was  to  be  established,  and  entrusted  with  such 
powers  as  might  enable  them  to  see  all  their  orders  executed  with  im- 
partial justice. 

2.  NotificfitioH. — The  ignorant  old  women  employed  as  searchers  were 
to  be  replaced  by  understanding  and  diligent  men,   who  were  to  report 
cases  immediately  to  the  Council  of  Health. 

3.  Isolation. — Physicians  were  at  once  to  be  despatched  to  visit  the 
suspected  cases,  and  when  the  suspicion  of  plague  was  confirmed,  all  the 
families  in  which  the  sickness  occurred  were  to  be  isolated.     The  sick 
were  to  be  separated  from  the  sound,  and  isolation  houses  to  be  provided 
three  or  four  miles  out  of  the  town. 

The  removal  of  the  sick  was  to  be  made  at  night,  so  as  to  avoid  the 
danger  of  spreading  infection,  and  all  possible  care  was  to  be  taken  to 
provide  such  means  of  conveyance  for  the  sick  that  they  might  receive  no 
injury.  The  poor  were  to  be  isolated  in  houses  provided  for  the  purpose, 
but  the  rich  were  to  be  allowed  to  be  in  their  own  homes  provided  that 
care  was  taken  to  separate  the  healthy  from  the  sick,  and  no  pains  were 
to  be  spared  to  provide  clean  and  airy  apartments.  All  expenses  were  to 
be  paid  by  the  public,  and  a  reward  was  to  be  given  to  the  person  who 
made  the  first  discovery  of  infection  in  any  place. 

Mead  further  pointed  out  that  general  sanitation  must  be  carefully 
attended  to.  Officers  were  to  see  that  the  streets  were  washed  and  kept 
clean  from  filth,  carrion,  and  all  manner  of  nuisances.  Beggars  and  idle 
persons  were  to  be  taken  up,  and  such  miserable  objects  as  were  fit 
neither  for  the  hospitals  nor  for  the  workhouses,  were  to  be  provided  for 
in  an  establishment  for  incurables.  Houses  also  were  to  be  kept  clean, 
and  sulphur  was  to  be  used  as  a  disinfectant. 

After  centuries  of  experience  we  have  learnt  that  the  necessary 
conditions  for  avoiding  the  plague  are  more  accurate  knowledge  on 
the  part  of  the  profession  and  the  public  of  the  way  in  which  the 
disease  spreads,  and  the  adoption  of  sanitary  precautions,  which  must 
include  personal  cleanliness,  sanitary  dwellings,  absence  of  overcrowd- 
ing, immediate  notification,  prompt  separation  of  the  sick  from  the 
healthy,  disinfection  of  infected  dwellings,  destruction  of  infected 
clothing,  and  extra-mural  burial  or,  better  still,  cremation.  It  was 
because  the  very  reverse  of  these  sanitary  conditions  existed  that 
the  virus  of  the  plague  found  a  suitable  environment  in  Old  London 
and  in  recent  times  in  Hong- Kong. 

RELAPSING  FEVER. 

Relapsing  or  famine  fever  is  a  contagious  disease  producing  a 
slate  of  high  fever  lasting  about  seven  days,  followed  by  apparent 

"  17 


258 


INFECTIVE    DISEASES. 


recovery,  and  in  about  fourteen  days  by  another  attack  of  fever, 
which  may  be  repeated  after  another  week. 

Starvation,  in  association  with  overcrowding  and  filth,  is  intimately 
connected  with  the  causation  of  the  disease.  The  subjects  of  the 
disease  contaminate  the  air  around  them,  and  the  virus  is  principally 
conveyed  by  tramps  and  dirty  people. 

Obermeier  discovered  spirilla  in  the  blood  during  the  paroxysms 
of  fever.  The  constant  occurrence  of  the  spirillum  in  relapsing 
fever,  and  the  fact  of  its  not  being  found  in  any  other  conditions, 
render  it  very  probable  that  it  is  the  cause  of  the  disease. 


D  N\ 


m 


FIG.  124.— SPIRILLUM  OBERMEIERI  IN  BLOOD  OF  MONKEY  INOCULATED  WITH 
SPIRILLA  AFTER  REMOVAL  OF  THE  SPLEEN  (SOUDAKEWITCH). 

Spirillum  Obermeieri  (Spirochceta  Obermeieri,  Oohn).— 
Threads  similar  to  the  Spirillum  plicatile.  In  length  they  are  mostly 
16  to  40  yu-,  with  regular  screw-curves.  They  move  very  rapidly, 
and  exhibit  peculiar  wave-like  undulations.  They  are  absent  from 
the  blood  during  the  non-febrile  intervals,  but  are  found  in  the 
interior  of  leucocytes  in  the  spleen.  In  blood  serum  and  50  per 
cent,  salt  solution,  they  preserve  their  movements.  In  cover-glass 
preparations  they  are  readily  stained  by  any  of  the  aniline  dyes, 
and  in  sections,  by  preference,  with  Bismarck  brown.  They  are  not 


TYPHUS   FEVER. — YELLOW   FEVER.  259 

found  in  the  urine,  sweat,  or  saliva.  They  have  not  been  cultivated 
artificially  on  any  nutrient  media.  Monkeys  have  been  successfully 
inoculated  with  blood  containing  the  spirilla  by  Koch,  Carter  and 
Soudakewitch ;  and  Koch  found  the  spirilla  in  the  vessels  of  the 
brain,  liver,  and  kidneys,  after  death.  According  to  Soudakewitch 
a  fatal  result  is  produced  in  monkeys  if  the  spleen  is  removed, 
and  the  spirilla  are  found  in  great  numbers  in  the  blood ;  but  if 
the  spleen  is  not  excised  the  spirilla  rapidly  disappear,  and  recovery 
follows.  Miinch  and  Motschutkowsky  transferred  blood  containing 
the  spirilla  to  healthy  persons,  and  produced  typical  relapsing  fever. 

TYPHUS  FEVER. 

Typhus  fever  is  a  highly  contagious  disease,  which  lasts  for  two 
or  three  weeks,  and  produces  a  measly  eruption.  Like  the  plague, 
it  is  intimately  associated  with  overcrowding  and  filth,  and  is  liable  to 
occur  where  these  conditions  exist  in  cities,  in  armies,  and  in  prisons. 
The  virus  produces  profound  changes  in  the  blood,  and  after  death 
the  internal  organs  are  found  to  be  congested,  especially  the  lungs, 
which  are  very  friable.  The  spleen  is  softened  and  often  enlarged, 
and  the  blood  is  dark  and  imperfectly  coagulated. 

The  virus  is  dissipated  by  fresh  air.  It  is  given  off  by  the 
breath  of  patients,  and  possibly  from  the  skin.  It  clings  to  the 
clothes  of  patients,  and  the  disease  may  be  conveyed  by  their  agency. 
One  attack,  as  a  rule,  confers  immunity.  Some  persons  are  naturally 
insusceptible,  failing  to  contract  the  disease  though  daily  exposed  to 
it.  Hlava  has  described  a  bacterium  which  he  believes  to  be  the 
specific  micro-organism.  Thoinot  and  Calmette  found  the  same 
bacterium  with  others,  but  there  was  no  particular  micro-organism 
constantly  present.  There  can  be  little  doubt  that  the  nature  of 
the  contagium  is  unknown. 

Stamping-out  System. — Sanitary  precautions,  and  especially 
the  operation  of  the  Public  Health  Acts  in  relation  to  lodging- 
houses,  prisons,  and  the  better  housing  of  the  working  classes,  have 
been  instrumental  in  almost  completely  stamping  out  the  disease  in 
this  country. 

YELLOW  FEVER. 

Yellow  fever  is  a  disease  of  tropical  climates,  characterised  by 
abdominal  tenderness,  hsemorrhagic  vomiting  (black- vomit),  and 
jaundice.  The  disease  may  end  fatally,  or  recovery  occur  in  about 
two  or  three  weeks.  It  is  especially  prevalent  in  the  West  Indies 
and  in  parts  of  North  and  South  America. 


260  INFECTIVE    DISEASES. 

The  virus  may  be  conveyed  by  infected  ships,  and  has  in  this 
way  made  its  appearance  at  British  and  French  seaport  towns. 
The  disease  is  generally  believed  to  be  contagious,  but  the  source 
of  the  virus  is  not  known.  According  to  Sternberg  the  virus  is  not 
conveyed  by  water,  but  spreads  where  there  is  overcrowding  and 
filth. 

Bacteria  in  Yellow  Fever. — Freire  asserts  that  there  is  a 
specific  micrococcus  in  yellow  fever  which  ca,n  be  grown  on  all 
ordinary  nutrient  media,  and  that  cultures  can  be  used  for  protective 
inoculation  with  satisfactory  results.  Carmonay  Yalle  also  claims  to 
have  discovered  the  contagium ;  but  Sternberg,  who  has  carried  on 
investigations  extending  over  several  years,  maintains  that  there  is  no 
characteristic  micro-organism  present  in  the  blood  or  in  the  tissues 
after  death.  Aerobic  and  anaerobic  cultures  were  made  from  the 
blood,  liver,  kidney,  urine,  stomach,  and  intestines.  The  liver  was 
found  to  contain  after  death  a  number  of  bacilli,  most  frequently 
Bacillus  coli  communis  and  Bacillus  cadaveris.  Blood  or  fresh 
liver  does  not  produce  any  disease  in  rabbits  or  guinea-pigs,  but 
liver  tissue  kept  for  forty-eight  hours  and  inoculated  subcutaneously 
in  guinea-pigs  is  extremely  pathogenic.  Similar  results  occur  after 
inoculation  of  healthy  liver  which  has  been  kept  in  the  same  way. 
We  may  conclude  from  these  experiments  that  the  nature  of  the 
contagium  is  unknown. 

Stamping-out  System. — Sternberg  states  that  there  are  many 
facts  relating  to  the  origin  and  extension  of  yellow  fever  epidemics 
which  support  the  theory  that  the  virus  is  present  in  the  evacua- 
tions, and  that  accumulations  of  fsecal  matter  and  of  organic 
material  of  animal  origin  furnish  in  certain  climates  a  suitable  soil 
for  the  development  of  the  contagium.  According  to  this  view  the 
evacuations  should  be  thoroughly  disinfected,  and  with  other  sanitary 
precautions  and  efficient  quarantine  at  seaports,  the  disease  may 
be  stamped  out,  and  the  danger  of  importation  from  the  natural 
home  of  the  disease  reduce:!  to  a  minimum. 


CHAPTER  XIX. 

SCARLET   FEVER. — MEASLES. 

SCARLET  FEVER. 

SCARLET  FEVER  is  a  highly  contagious  disease  peculiar  to  man.  It 
produces  inflammation  of  the  tonsils  and  adjoining  parts,  fever,  and 
a  general  punctiform  eruption.  The  period  of  incubation  is  about 
a  week,  and  the  rash  usually  appears  on  the  second  day.  In  some 
cases  the  disease  manifests  itself  in  an  extremely  mild  form,  known 
as  latent  scarlet  fever,  in  which  there  is  only  a  slight  febrile  attack, 
or  a  mild  sore  throat,  with  very  little  or  no  rash.  Many  cases 
would  not  be  recognisable  as  such  if  they  were  not  capable  of 
conveying  scarlet  fever,  or  unless  other  cases  followed  or  occurred 
simultaneously  which  were  undoubtedly  typical  cases  of  the  disease. 
The  occurrence  of  such  cases  in  the  early  history  of  an  epidemic 
often  causes  the  greatest  difficulty  in  tracing  the  origin  of  the 
outbreak,  and  indeed  in  some  cases  renders  it  quite  impossible  to 
do  so. 

The  virus  is  given  off  by  the  skin,  in  desquamation,  and  possibly 
by  the  urine.  It  maintains  its  vitality  in  clothing  for  months,  and 
sometimes  longer.  It  may  also  be  conveyed  by  the  hands  of  the 
physician  to  women  during  parturition.  The  disease  may  be 
transferred  by  subcutaneously  inoculating  persons,  who  have  not 
previously  contracted  scarlet  fever,  with  virus  obtained  by  puncturing 
the  eruption  on  the  skin. 

After  death  the  internal  organs  appear  to  the  naked  eye  more  or 
less  healthy.  The  liver  is  soft,  the  kidneys  are  congested,  the  ileum 
is  inflamed,  and  Peyer's  patches  enlarged  and  congested;  but  these 
conditions  are  also  produced  by  other  causes.  There  are  inflam- 
matory changes  in  the  lymphatic  follicles  of  the  tonsils,  and  the 
larynx  and  trachea.  Other  morbid  lesions,  especially  in  the  kidneys, 
are  associated  with  the  sequelae  and  complications,  and  though 
commonly  occurring  in  scarlet  fever  are  also  found  in  other  diseases. 

261 


262  INFECTIVE   DISEASES. 

These  changes  appear  to  be  clue  to  the  poison  which  is  in  the  blood, 
and  is  excreted  by  the  kidneys.  The  epithelium  is  in  a  state  of 
cloudy  swelling,  a  condition  found  in  other  febrile  diseases  and  in 
septic  poisoning. 

Bacteria  in  Scarlet  Fever. — The  occurrence  of  micro-organ- 
isms in  cases  of  scarlet  fever  has  been  observed  by  several  investi- 
gators— Coze  and  Feltz,  Crooke,  Loftier,  Babes,  Heubner  and  Bahrdt, 
and  notably  by  Frankel  and  Freudenberg,  and  more  recently  by 
Klein,  the  author,  Raskin,  and  others. 

Coze  and  Feltz  found  cocci  in  the  blood,  and  Crooke,  in  cases  of 
scarlet  fever  with  severely  affected  throat,  found  bacilli,  cocci,  and 
streptococci  in  the  organs  of  the  throat,  and  cocci  in  the  internal 
organs.  Crooke  left  it  an  open  question  whether  these  cocci  were 
the  specific  organisms  of  scarlet  fever,  or  were  to  be  regarded  as 
diphtheritic  or  septic  associates.  He  inclined,  for  clinical  reasons, 
to  the  latter  view. 

Loffler,  in  cases  of  scarlatinal  diphtheria,  found  the  same  chain - 
forming  micrococcus  which  he  had  found  in  typical  diphtheria. 

Babes  was  able  constantly  to  prove  the  presence  of  a  strepto- 
coccus in  inflammatory  products  secondary  to  scarlatina. 

Heubner  and  Bahrdt,  in  a  fatal  case  of  scarlet  fever  in  a  boy, 
complicated  with  suppuration  of  the  finger  and  knee-joints,  and 
with  pericarditis,  found  a  streptococcus  identical  in  form  with  Strepto- 
coccus pyogenes,  but  cultivations  were  not  made.  The  secondary 
infection  started  from  diphtheritically  affected  tonsils,  which  were 
followed  by  retro-pharyngeal  abscesses. 

Frankel  and  Freudenberg  examined,  for  micro-organisms,  three 
cases  of  scarlatina  with  well-marked  affection  of  the  throat.  In  all 
three  cases  they  obtained  cultivations  of  cocci  from  the  submaxillary 
lymphatic  glands,  spleen,  liver,  and  kidney.  These  cocci  could  not 
be  distinguished  from  Streptococcus  pyogenes  derived  from  pus,  nor 
from  the  undoubtedly  identical  streptococcus  which  one  of  them 
(A.  Frankel)  had  repeatedly  cultivated  in  large  numbers  from 
puerperal  affections.  In  two  of  the  cases  Streptococcus  pyogenes 
was  the  only  organism  present,  and  in  all  three  cases  it  was  far  in 
excess  of  other  colonies  which  developed.  The  organisms  were  also 
found  in  sections  of  the  organs  by  microscopical  examination. 
Frankel  and  Freudenberg  could  in  no  way  distinguish  the  strepto- 
coccus in  scarlatina  from  the  streptococcus  in  pyaemia  and  septi- 
caemia. The  identity  of  this  streptococcus  with  Streptococcus  pyogenes 
and  Streptococcus  puerperalis  was  established  by  comparison  of  their 
macroscopical  and  microscopical  appearances  in  cultivations  on 


SCARLET  FEVER.  263 

nutrient  agar-agar,  nutrient  gelatine,  and  in  broth,  both  at  the 
ordinary  and  at  higher  temperatures,  and  also  by  experiments  on 
animals.  They  concluded  that  it  could  be  stated  with  certainty 
that  the  organisms  in  question  did  not  stand  in  causal  relation  to 
scarlet  fever.  They  considered  that  special  methods  of  microscopical 
and  biological  research  were  apparently  needed  for  demonstrating 
the  true  scarlet  fever  contagium,  which  probably  was  especially 
present  in  the  skin.  They  considered  that  the  presence  of  the 


a.  b.  c. 

FlG.    125.— PURE-CULTIVATIONS  OF   STREPTOCOCCUS   PYOGENES. 

(a)  On  the  surface  of  nutrient  gelatine ;  (b)  In  the  depth  of  nutrient  gelatine  : 
(c)  On  the  surface  of  nutrient  agar. 

streptococcus  was  due  to  a  secondary  infection,  to  which  the  door 
was  opened  by  the  lesions  of  the  throat — a  view  which  was  sup- 
ported by  the  fact  that  the  organisms  were  found  in  submaxillary 
lymphatic  glands.  They  preferred  to  use  the  term  "  secondary  " 
to  "  complicated  "  or  "  combined  "  infection,  because  this  expresses 
the  fact  that  by  the  effect  of  the  scarlatinal  virus  the  soil  is 
rendered  suitable  for  this  ubiquitous  microbe  when  it  has  once 
gained  an  entrance. 

This  streptococcus  was  found  by  Klein  in  five  out  of  eleven  cases 


INFECTIVE    DISEASES. 


of  scarlet  fever  in  man,  twice  in  association  with  certain  other 
micro-organisms,  and  three  times  alone.  The  micro-organisms  were 
isolated  by  inoculating  tubes  of  nutrient  gelatine,  solidified  obliquely, 
by  streaking  the  surface  with  blood  taken  from  the  finger,  the  arm, 
or  the  heart  after  death.  Those  cases  from  which  the  organism  was 
obtained  were  all  cases  with  ulcerated  throat,  and  the  culture 
experiments,  from  the  living  patient,  were  made  on  or  about  the  day 
at  which  the  temperature  was  at  its  maximum. 


Name  of  Patient. 

Condition  of 
Tonsils. 

Source            Micro-Organisms 
Blood.                    Isolated' 

Death  or 
Recovery. 

1 

L  F  . 

Severely 

Finger 

Streptococcus 

Ultimately 

aged  5 

ulcerated 

recovered. 

fStaphylococcus 

] 

2 

K  F  , 

aged  2 

Much 
ulcerated 

.      " 

pyogenes  aureus 
•(  Liquefying    micro- 
coccus 

1  Died  of 
f     pygemia. 

^Streptococcus 

J 

3 
4 

H  —  L  —  , 

aged  8 
(a  woman), 

Ulcerated 
Much 

Arm 

J  Staphylococcus 
(Streptococcus 
None 

[•Recovered. 
[Not  stated.] 

aged  40 

ulcerated 

5 

(a  girl), 

55 

•5 

5J 

aged  19 

6 

B  M  , 

Ulcerated 

Finger  i        Streptococcus 

Recovered. 

aged  15 

7 

E  W  . 

Much 

„                     None 

[Not  stated.] 

aged  22 

ulcerated 

8 

R  H  , 

Ulcerated 

• 

;» 

aged  8 

9 

F  G  , 

M 

Heart          Streptococcus 

Died. 

aged  2J 

10 

E          F 

— 

„                     None 

,, 

aged  3 

11 

E  B  , 

Advanced 

„       | 

5J 

aged  20  months 

ulceration 

i 

Klein  regards  this  streptococcus  as  the  actual  cause  of  scarlet 
fever  in  man. 

The  author,  Raskin,  Holmes,  and  others  who  have  investigated  this 
subject  agree  with  the  conclusions  of  Fraiikel  and  Freudenberg.  The 
author  is  convinced  that  the  streptococci  in  suppuration,  puerperal 
septicaemia,  pyaemia,  and  septicaemia,  and  in  certain  cases  of  measles, 
scarlatina,  and  diphtheria,  are  identical ;  and  from  overwhelming 
evidence  we  are  justified  in  concluding  that — (1)  The  nature  of 
the  contagium  of  scarlet  fever  is  unknown.  (2)  The  streptococcus 
regarded  by  Klein  as  the  contagium  is  the  Streptococcus  pyogenes. 


MILK-SCARLATINA.  265 

(3)  This  streptococcus  is  found,  sometimes  in  company  with  Staphylo- 
coccus  pyogeiies  aureus,  as  a  secondary  result  in  scarlet  fever  and 
many  other  diseases,  and  its  exact  relation  to  scarlet  fever  and  its 
identity  with  the  streptococcus  from  pus  and  puerperal  fever,  were 
definitely  established  in  1885  by  Frankel  and  Freudenberg. 

MILK-  SCARLATINA. 

It  would  not  be  necessary  to  say  anything  further  on  the  etiology 
of  scarlet  fever  if  the  generally  accepted  belief,  that  scarlet  fever  is 
a  disease  peculiar  to  man,  were  accepted  by  the  Medical  Department 
of  the  Local  Government  Board;  but  the  theory  is  officially  held 
that  scarlet  fever  is  in  its  origin  a  disease  of  cows.  Bovine  scarlatina 
is  supposed  to  be  an  eruptive  disease  of  the  teats,  and  it  is  maintained 
that  the  virus,  by  contaminating  the  milk,  produces  scarlet  fever  in 
the  human  subject.  As  this  theory  is  very  naturally  accepted  by 
many  medical  officers  of  health,  and  is  mentioned  in  English  medical 
text-books,  it  will  be  necessary  to  discuss  this  question  in  considerable 
detail,  and  especially  as  these  opinions  were  promulgated  in  this 
country  with  official  support,  and  have  since  been  proved  to  be 
erroneous. 

The  theory  of  the  origin  of  the  exanthemata  in  diseases  of  the 
lower  animals  is  a  very  old  one.  The  Arabians  imagined  that  small- 
pox arose  from  the  camel.  Jenner  adopted  a  similar  theory,  and 
expressed  his  belief  that  small-pox  originated  in  the  horse,  being 
generated  by  horses  suffering  with  "  greasy "  hocks.  Thus  Jenner 
wrote  :  "May  not  accidental  circumstances  have  again  and  again 
arisen,  still  working  new  changes  upon  it,  until  it  has  acquired 
the  contagious  and  malignant  form  under  which  we  now  com- 
monly see  it,  making  its  devastations  among  us  ?  and  from  a 
consideration  of  the  change  which  the  infectious  matter  undergoes 
from  producing  a  disease  in  the  cow,  may  we  not  conceive  that 
many  contagious  diseases  now  prevalent  amongst  us  may  owe  their 
present  appearance,  not  to  a  simple,  but  a  compound  origin  ?  For 
example,  is  it  difficult  to  imagine  that  measles,  scarlet  fever,  ulcerated 
sore  throats,  and  spotted  skin,  all  spring  from  the  same  source, 
assuming  some  variety  in  their  forms  according  to  the  nature  of 
their  new  combinations?"  Baron  informs  us  that  tliis  idea  was 
prevalent  in  Jeimer's  mind  as  early  as  1787.  It  is  related  that  in 
that  year  he  accompanied  his  nephew,  George  Jenner,  into  a  stable 
to  look  at  a  horse  with  diseased  heels,  and,  pointing  to  them,  he 
it-marked  :  "  There  is  the  source  of  small-pox.  I  have  much  to  say 


266  INFECTIVE    DISEASES. 

on  that  subject  which  I  hope  in  due  time  to  give  to  the  world." 
And  again  in  1794,  when  writing  in  connection  with  this  subject, 
he  adds  :  "  Domestication  of  animals  has  certainly  provided  a  prolific 
source  of  diseases  among  man." 

Jenner's  views  were  found  to  be  incorrect,  and  it  was  shown  by 
Loy  and  others  that  the  grease  bears  no  relation  to  cow-pox,  and 
it  is  now  known  that  Jenner  mistook  horse-pox  for  the  disease 
known  as  the  grease.  No  one  at  the  present  day  supports  Jenner's 
theory  of  small-pox  in  man  arising  from  any  disease  of  the  horse. 
Indeed,  the  origin  of  small-pox  from  a  disease  of  the  horse  was  not 
upheld  even  by  Jenner's  pupil  and  nephew,  Henry  Jenner.  The 
latter  promulgated  the  idea  that  small- pox  originated  from  the  cow. 
He  believed  that  small-pox,  in  fact,  was  cow-pox  intensified  in  its 
virulence  by  being  passed  through  man.  He  thus  expressed  himself  • 
"  Nor  may  it,  perhaps,  be  too  hypothetical  to  suppose  that  the 
cow-pox  may  possibly  be  the  small-pox  in  its  original  unadulterated 
state,  before  it  became  contaminated  by  passing  through  the  impure 
and  scrofulous  habits  of  human  constitutions."  The  theory  of  the 
origin,  in  animals,  of  human  febrile  diseases  was,  later,  advocated  by 
Copland,  who  stated,  firstly,  that  scarlet  fever  in  man  was  originally 
a  disease  of  the  horse,  and  that  it  formerly  occurred,  and  had  recently 
occurred,  epidemically  as  an  epizootic  among  horses ;  secondly,  that 
it  was  communicated  in  comparatively  modern  times  from  horses  to 
man ;  thirdly,  that  it  might  be,  and  had  been,  communicated  to  the 
dog.  But  this  opinion  has  not  been  accepted,  for  the  disease  called 
scarlatina  in  the  horse  is  a  lion- infectious  disease,  generally  attack- 
ing but  one  or  two  horses  in  a  large  stud.  •  It  neither  spreads  by 
contagion  nor  infection  ;  and  Williams  states  that  it  is  impossible  to 
transmit  it  from  the  horse  to  any  other  animal,  and  that  many  cases 
of  the  so-called  scarlatina  of  the  horse  are  in  reality  identical  with 
purpura. 

The  theory  was  again  revived,  but  in  another  form,  and  has 
been  adopted  by  the  Medical  Department  of  the  Local  Government 
Board.  Owing  to  failure  in  tracing,  in  some  cases  of  milk  scarla- 
tina, the  contamination  of  the  milk  from  a  human  source,  the 
theory  was  started  that  in  such  cases  the  disease  is  derived  from 
the  cow — that,  in  other  words,  there  is  a  disease,  scarlet  fever 
in  the  cow,  which  is  responsible  for  outbreaks  of  scarlet  fever  in 
man. 

In  1882  an  epidemic  of  scarlatina  in  St.  Giles  and  St.  Pancras 
was  investigated  by  Mr.  W.  H.  Power  for  the  Board.  The  disease 
was  distributed  with  a  milk  supply  from  a  Surrey  farm.  In  this  case 


MILK-SCARLATINA.  267 

two  facts  were  ascertained  :  the  one,  that  a  cow  recently  come  into 
milk  had  been  suffering  from  some  ailment  from  the  time  of  her 
parturition,  of  which  loss  of  hair  in  patches  was  the  most  con- 
spicuous manifestation  ;  the  other,  that  there  existed  no  discoverable 
means  by  which  the  milk  could  have  received  infective  quality  from 
the  human  subject. 

In  1885  an  outbreak  of  scarlet  fever  occurred  in  Marylebone  in 
connection  with  milk  from  a  farm  at  Hendoii,  and  again  Power 
failed  to  establish  infection  from  any  human  source  in  any  commonly 
accepted  way — such,  for  example,  as  handling  of  milk,  or  milk  utensils, 
by  persons  carrying  scarlatina  infection.  But  on  examining  the 
cows  with  a  view  to  ascertain  any  new  condition  pertaining  to 
them,  it  came  to  light  during  the  inquiry  that  some  of  them, 
which  had  recently  been  introduced  from  Derbyshire,  were  suffering 
from  a  vesicular  disease  of  the  teats. 

At  this  stage  Klein  became  associated  with  Power  in  the 
inquiry;  and  their  belief  in  the  existence  of  a  disease  among  the 
cows  on  the  farm  capable  of  producing  scarlatina  among  human 
consumers  of  the  cow's  milk,  became  unreserved.  Klein  took  away 
with  him  samples  of  milk,  contents  of  vesicles,  and  discharges 
from  ulcers,  and  afterwards  two  of  the  cows  were  purchased  and 
kept  under  observation. 

Dr.  Cameron  of  Hendon  has  given  a  detailed  description  of  the 
clinical  history  of  this  disease.  He  expressed  his  belief  that  it  was 
a  specific  disease  capable  of  being  communicated  to  healthy  cows 
by  direct  inoculation  of  the  teats  with  virus  conveyed  by  the  milker 
from  a  diseased  animal. 

The  condition  of  the  teats  is  described  as  follows  :  The  teats 
became  enlarged,  swollen  to  nearly  twice  their  natural  size,  and 
cedematotis.  On  handling  them  there  was  no  feeling  of  induration. 
Vesicles  appeared  on  the  swollen  teats  and  upon  the  udder  between 
or  near  the  teats.  These  varied  in  number  from  two  to  four  on  a 
teat,  and  in  size  from  a  pea  to  a  horsebean.  The  vesicle  contained 
a  clear  fluid.  The  vesicles  were  rubbed  and  broken  in  milking,  and 
left  raw  sores,  sometimes  red,  in  other  cases  pale  in  colour,  with 
raised,  ulcerated  edges.  Sometimes  a  few  accessory  vesicles  formed 
around  the  margin  of  these  ulcerated  sores.  After  the  rupture  of 
the  vesicle  a  brown  scab  formed,  which  might  remain  attached  for 
five  or  six  weeks,  or  fall  off  in  ten  days  or  a  fortnight,  a  smaller 
one  forming ; afterwards.  A  thin,  watery  fluid  exuded  from  un.lrr 
the  scab,  and  the  sore  ultimately  healed. 

Cameron  examined  the  teats  of  several  cows  five  or  six  weeks 


268  INFECTIVE    DISEASES. 

after  they  were  attacked.  The  scabs  then  varied  in  size  from  a 
shilling  to  a  florin ;  they  were  about  one-eighth  of  an  inch  thick  in 
the  centre,  thinning  off  towards  the  edges. 

Some  of  the  cows  were  also  suffering  from  an  eruption  on  the 
rump  and  hind  quarter,  consisting  of  patches  of  eczematous  crusts. 
When  a  crust  was  picked  off,  the  hair  came  off  with  it,  exposing  a 
raw,  moist  sore,  the  crusts  and  sores  looking  exactly  like  eczematous 
scabs  and  sores  ;  but  this  condition  corresponds  in  description  with 
eczema,  the  result  of  ringworm  which  is  very  common  in  young 
stock. 

In  addition  to  his  own  observations,  Cameron  obtained  infor- 
mation from  the  farmers,  and  others  familiar  with  cows,  who 
thought  they  recognised  in  the  disease  at  the  farm  one  stage  of  a 
disease  which  they  were  able  to  describe.  Cameron  thus  gives  an 
account  of  what  he  and  his  informants  together  would  regard  as 
a  connected  clinical  history  of  the  disease. 

He  did  not  see  the  earlier  symptoms,  and  hence  these  were 
of  necessity  learnt  from  other  persons.  The  account,  therefore, 
of  these  symptoms  was  to  be  held  liable  to  future  correction  or 
modification. 

Cameron  stated  that  he  learnt  this  disease  was  capable  of 
being  communicated  to  milkers  by  inoculation  with  virus  from  the 
vesicles  on  the  teats,  though  the  milkers  on  the  Hen  don  farm 
escaped.  "  A  trusty  informant  received  the  virus  into  a  recent 
scratch  on  the  forefinger  while  milking  a  diseased  cow.  General 
weakness,  malaise,  and  loss  of  appetite  resulted,  and  after  about 
four  or  five  days  a  vesicle  or  small  blister  appeared  on  the  finger. 
This  broke,  and  several  others  formed  on  the  back  of  the  hand. 
The  whole  hand  and  fingers  became  swollen  and  inflamed,  the 
inflammation  extending  in  broad  lines  as  far  as  the  elbow.  The 
general  disturbance  lasted  a  fortnight." 

In  the  course  of  the  inquiry,  Cameron  adds  that  it  was 
strongly  asserted  by  several  people,  who  examined  the  cows,  that 
they  were  suffering  from  cow-pox.  He,  however,  dismissed  the 
diagnosis  of  cow-pox  on  the  ground  that  no  papule  had  been 
observed  or  subsequent  formation  of  pustule,  areola,  or  pitting,  and 
because  the  vesicles  were  not  umbilicated.  These  reasons  given 
for  dismissing  the  diagnosis  of  cow-pox  at  Hendon  were  totally 
inadequate ;  a  comparison  having  been  made  between  the  characters 
of  the  eruption  of  vaccinia  as  it  appears  on  an  infant's^arm,  instead 
of  the  eruption  of  the  natural  or  so-called  spontaneous  disease  011 
the  teats  of  the  cow. 


MILK-SCARLATINA. 

Klein  stated  that  on  the  teats  and  udders  of  two  cows  which 
he  investigated  there  were  several  flat  irregular  ulcers,  varying  in 
diameter  from  one-quarter  to  three-quarters  of  an  inch.  Some 
ulcers  were  more  or  less  circular,  others  extended  in  a  longitudinal 
direction  on  the  teat.  The  ulcers  were  covered  with  a  brownish  or 
reddish-brown  scab.  The  animals  looked  thin,  but  not  strikingly  so. 
In  feeding  capacity,  milking  power,  arid  body  temperature  there 
was  nothing  abnormal. 

Four  calves  were  inoculated  in  the  corium  of  the  groin  and  the 
inside  of  the  ear,  with  scrapings  from  the  ulcers  after  removal  of 
the  crusts.  In  one,  which  may  be  taken  as  an  example  of  the  result 
obtained,  there  was  vesiculation  at  the  margin  of  the  spot  inoculated, 
and  in  the  centre  the  commencement  of  the  formation  of  a  crust. 
On  the  seventh  day  each  sore  on  the  ear  had  enlarged  to  about 
half  an  inch  in  breadth,  and  was  covered  in  its  whole  extent  by 
a  brownish  crust.  On  the  eighteenth  day  they  had  all  healed  up 
and  become  converted  into  flat  scars. 

To  search  for  micro-organisms,  Klein  removed  the  crust 
from  an  ulcer  on  the  teat,  scraped  off  the  most  superficial  layer, 
squeezed  the  ulcer,  and  made  cover-glass  preparations.  Tubes  of 
nutrient  gelatine  and  nutrient  agar-agar  were  also  inoculated,  and 
a  streptococcus  was  isolated  which  in  morphological  and  cultural 
characters  agreed  with  those  of  Streptococcus  pyogeues. 

Two  calves  were  inoculated  in  the  groin  with  the  cultivated 
micro-organism.  One  calf  died  in  twenty-seven  days.  At  the 
necropsy  there  were  found  peritonitis,  and  hsemorrhagic  spots  on 
the  omentum ;  the  liver,  kidneys,  and  lungs  were  congested,  and 
there  were  petechise  under  the  pleura,  and  pericarditis.  The  second 
calf  was  killed,  and  at  the  necropsy  the  lungs  and  kidneys  were 
congested,  and  there  were  hsemorrhagic  patches  on  the  spleen. 

In  these  cases,  the  post-mortem  appearances  and  anatomical 
features  recalled  to  Klein  the  lesions  of  scarlatina.  In  the 
kidney,  for  example,  the  cortex  was  congested,  and  there  were 
haemorrhages,  glomerulo-nephritis,  and  granular  or  opaque  swelling 
of  the  epithelial  cells  and  infiltration  with  round  cells.  From  the 
blood  of  the  heart  the  streptococcus,  which  had  been  used  in  the 
inoculation,  was  recovered.  In  view  of  this  evidence  it  was  con- 
cluded that  the  streptococcus  was  the  virus  of  the  cow  disease,  and 
that  it  produced  in  calves  a  disease  very  closely  resembling  that  of 
scarlatina  in  man. 

Two  of  the  cows  selected  from  the  Hendon  farm  were  killed, 
and  it  was  observed  in  one  that  the  lungs  were  congested,  and 


270  INFECTIVE   DISEASES. 

that  there  were  numerous  adhesions  by  recent  soft  lymph  between 
the  lower  lobes  of  the  lung  and  the  costal  pleura.  In  the 
liver  there  were  several  reddish  streaks  and  patches.  The  spleen 
and  kidneys,  with  the  exception  of  slight  congestion,  appeared 
normal. 

Sections  of  the  kidney  showed  well-marked  glomerulo- nephritis 
and  infiltration  of  the  sheath  of  the  cortical  arterioles  with  numerous 
round  cells.  The  epithelium  of  the  convoluted  tubules  was  swollen, 
opaque,  and  in  many  places  disintegrating. 

In  the  other  cow  there  was  great  congestion  of  the  lungs  and 
pleural  adhesions  ;  the  cortex  of  the  kidney  was  congested,  but  its 
medulla  was  pale. 

On  microscopic  examination  there  was  a  good  deal  of  round - 
celled  infiltration  in  the  walls  of  the  inf  undibula  and  bronchi  in  the 
lung,  and  round  the  arterioles  in  the  kidney. 

In  sections  of  the  ulcers  on  the  teats,  the  corium  was  found  to  be 
infiltrated  throughout  the  whole  extent  of  the  ulcer  with  round 
cells.  In  the  superficial  layers  of  the  stratum  Malpighi,  close  to 
the  stratum  lucidum,  as  also  in  the  stratum  lucidum  itself,  there 
were  numerous  cavities  of  different  sizes.  These  cavities  lay  side  by 
side,  the  most  superficial  ones  being  covered  by  the  stratum  lucidum, 
or  extending  between  the  layers  of  this  stratum. 

At  the  marginal  parts  the  cavities,  although  placed  side  by  side, 
were  well  separated  from  one  another  by  thicker  or  thinner 
trabeculse,  composed  of  epithelium,  while  at  or  near  the  centre  of 
the  ulcer  these  trabecula?  were  destroyed,  the  cavities  had  become 
confluent,  and  the  covering  layers  of  the  cuticle  having  here  also 
given  way,  their  contents  extended  on  to  the  free  surface  of  the 
ulcer.  In  short,  Klein  states  that  all  the  anatomical  details  of 
the  distribution  and  arrangement  of  these  cavities  recalled  vividly 
the  conditions  observed  in  the  vesicles  of  cow-pox.  Yet  as  a  result 
of  this  investigation  he  concluded  that  the  cow  disease  at  Heridon 
was  bovine  scarlatina,  and  that  towards  its  study  and  supervision 
every  effort  ought  to  be  directed  in  order  to  check  the  spread  of 
scarlet  fever  in  man. 

As  a  result  of  this  conclusion,  the  Board  of  Agriculture  resolved 
to  have  the  whole  subject  fully  investigated,  and  the  author  was 
directed  to  study  the  bacteriology  and  micro-pathology  of  this  disease 
and  to  report  thereon.  Professor  Axe  investigated  the  origin  of 
the  outbreak  of  the  disease  in  the  cows,  and  Professor  M'Fadyean 
carried  out  an  investigation  into  the  possibility  of  inoculating  cows 
with  the  virus  from  cases  of  scarlet  fever  in  man. 


MILK-SCARLATINA.  271 


THE  AUTHOR'S  INVESTIGATION. 

An  outbreak  of  an  eruptive  'disease  of  the  teats,  alleged  to  be 
identical  with  the  so-called  Hendon  cow  disease,  was  raging  in  some 
farms  in  Wiltshire.  In  this  case  every  facility  was  given  by  the 
owner  of  the  estate  for  a  thorough  investigation  into  the  disease. 
Not  only  were  animals  sent  from  the  farm  to  London,  but  the 
author  was  allowed  to  visit  the  farms,  to  inspect  all  the  infected 
animals,  and  to  make  every  investigation,  with  the  hearty  co- 
operation of  the  bailiff  of  the  farms,  and  the  voluntary  assistance 
of  the  head  cowmen  and  those  under  them.  Some  of  these  cowmen 
were  unusually  intelligent,  while  two  had  had  experience  of  cows 
for  more  than  half  a  century.  Thus,  there  was  not  only  every 
opportunity  for  studying  the  disease  on  the  lines  indicated  by 
Klein,  but  it  was  possible  by  repeated  visits  to  the  farms  to  enter 
into  the  clinical  history  of  the  disease  in  the  cow,  to  study  very  fully 
the  nature  of  the  disease  on  the  hands  of  the  milkers,  and  to  trace 
the  probable  mode  of  its  introduction  on  the  estate,  and  the  way  in 
which  it  spread  from  one  part  of  the  herd  to  another. 

Two  cows  were  sent  to  London  with  disease  of  the  teats  and  of 
the  udder  between  the  teats.  On  the  right  teats  of  one  there  were 
numerous  sores,  covered  with  crusts  varying  in  size  and  in  thickness, 
and  generally  fissured.  In  some  they  were  flat,  in  others  conical  ; 
some  were  with  difficulty  removed  with  forceps,  others  were  readily 
detached.  The  crusts  varied  in  colour  from  reddish-brown  to  very 
dark  brown  or  almost  black.  On  detaching  or  scraping  a  crust 
there  was  a  granulating  and  somewhat  indurated  base.  On  the 
right  anterior  teat  there  were  several  ulcers,  from  which  appa- 
rently the  thick  crusts  had  been  detached,  and  new  scabs  were 
forming.  On  the  left  posterior  teat  there  were  unusually  large, 
dark  brown,  or  blackish  crusts,  covering  a  very  extensive  are;i 
of  ulceration,  extending  over  the  whole  of  the  lower  third  of  the 
teat. 

In  the  other  cow  from  Wiltshire  there  was  the  same  disease  on 
the  teats,  but  not  in  such  a  severe  form.  The  sores  were  covered 
with  thick  crusts,  but  though  varying  in  size  they  were  more 
regular  in  form,  and  more  circumscribed. 

Having  entirely  removed  the  crusts  from  some  of  the  ulcers,  a 
number  of  inoculations  in  nutrient  gelatine  and  nutrient  agar-agar 
were  made  from  the  discharge,  and  cover-glass  preparations  were 
made  and  stained  in  the  ordinary  way.  Cultures  were  obtained  of 
the  organisms  commonly  found  in  pus. 


272  INFECTIVE    DISEASES. 

With  the  discharge  and  with  scrapings  from  the  ulcers  two 
calves  were  inoculated. 

Of  the  two  calves,  one  was  inoculated  by  scarification  in  both 
ears  ;  the  other,  a  small  calf,  was  scarified  in  the  left  ear.  Scrapings 
from  the  ulcers  were  rubbed  into  the  places  thus  prepared.  In 
addition,  in  the  small  calf  an  incision  was  made  through  the  corium 
in  the  left  groin,  scrapings  from  different  ulcers  on  the  teats  were 
well  rubbed  in  with  the  blade  of  the  scalpel,  and  a  portion  of  crust 
inserted  into  a  small  pocket  in  the  subcutaneous  tissue.  In  the  ears 
and  the  groin  there  were  positive  results.  In  the  large  brown  calf 
one  of  two  places  inoculated  in  the  right  ear  passed  through  the 
following  changes  :  On  the  third  day  there  was  apparent  vesiculation 
and  commencing  formation  of  crust.  From  day  to  day  the  crust 
thickened,  and  on  the  eighth  day  the  crust  was  at  its  height  and 
detached  at  its  edges.  By  removing  the  scab  an  ulcer  was  exposed  ; 
there  was  slight  inflammatory  thickening.  About  the  thirteenth 
day  the  ulcer  had  quite  healed. 

Very  similar  appearances  resulted  in  the  ear  of  the  smaller  calf. 
The  result  of  inoculation  in  the  groin  was  of  a  very  much  severer 
character.  In  the  course  of  two  or  three  days  the  incision  had 
apparently  commenced  to  heal  by  scabbing,  but  there  was  a  surround- 
ing area  which  was  inflamed,  and  painful  on  manipulation.  The 
inflammatory  thickening  which  resulted  continued  to  increase  around 
the  seat  of  inoculation,  and  the  thickening  could  be  felt  to  extend 
deeply  into  the  groin.  Suppuration  followed,  and  on  firm  pressure 
pus  welled  up  through  the  wound.  The  wound  then  showed  very 
little  disposition  to  heal,  and  the  calf  began  to  exhibit  marked 
constitutional  symptoms.  During  the  second  week  after  inoculation 
the  animal  became  very  dull,  and  was  reported  by  the  attendant  as 
refusing  to  feed.  Diarrhoea  supervened,  and  lasted  for  several  days, 
and  bloody  urine  was  passed.  The  calf  was  also  noticed  to  cough, 
and  the  cough  gradually  increased  in  severity.  Thirty-six  days  after 
the  date  of  inoculation  it  was  decided  to  kill  the  calf  and  examine 
the  condition  of  the  viscera.  The  appearances  which  were  found 
at  the  post-mortem  examination  were  as  follows  : — 

The  upper  and  middle  lobes  of  each  lung  were  adherent  to  the  walls 
of  the  chest  ;  there  was  congestion,  especially  of  the  middle  lobe,  and 
patches  of  recent  adherent  lymph.  Posterior  parts  of  the  upper  lobes  of 
both  lungs  were  completely  consolidated,  and  on  section  varied  in  colour 
from  brick-red  to  greyish -white.  The  interlobular  tissue  was  infiltrated 
with  inflammatory  products,  which  mapped  out  the  tissue  of  the  lung  in 
small  indurated  areas,  in  which  the  tissue  was  granular-looking  and  friable. 


THE  AUTHOR'S  INVESTIGATION.  273 

These  appearances  in  the  upper  lobes  were  due  to  septic  pleuro-pneumonia. 
They  closely  resembled,  and  were  supposed  to  be  due  to,  infectious 
pleuro-pneumonia.  They  were,  however,  found  identical  with  the  con- 
dition observed  in  septic  pleuro-pneumonia  in  calves,  and  the  disease  was 
not  conveyed  by  infection  to  other  animals  in  the  same  stall.  Scattered 
through  the  other  lobes  of  both  lungs  were  white,  mostly  firm,  nodules 
raised  above  the  level  of  the  surface  of  the  lung.  They  were  surrounded 
by  a  zone  of  congestion,  and  in  some  cases  sections  were  composed  of 
indurated,  in  others  of  friable,  lung  tissue.  In  the  posterior  part  of  the 
right  upper  lobe  there  was  a  recent  infarct.  The  bronchial  glands  at  the 
roots  of  each  lung  were  enlarged  to  two  or  three  times  their  natural  size, 
and  were  firm  and  hard  on  section.  The  parietal  surface  of  the  pericardium 
was  covered  with  recent  adherent  lymph.  The  visceral  surface  of  the 
pericardium  was  normal.  Along  the  external  surface  of  the  aorta  were 
chains  of  enlarged  lymphatic  glands  connected  by  dilated  lymphatic 
vessels.  These  glands  were  dark  red  or  purplish  in  colour,  from  haemor- 
rhage into  their  substance.  The  heart  was  normal,  and  the  endocardium 
not  stained.  There  were  chains  of  red  glands  on  the  oesophagus  similar 
to  those  along  the  aorta.  The  appearance  of  the  mesenteric  glands  was 
very  striking.  The  mesentery,  along  the  lymphatic  vessels,  was  dotted 
with  glands,  varying  in  size  from  a  large  shot  to  a  pea,  which  were  deep 
red  or  prune-coloured.  In  addition,  there  were  here  and  there  enlarged 
glands  without  haemorrhage  into  their  substance,  and  greyish  in  colour. 
There  were  scattered  petechiae  on  the  spleen.  The  kidneys  were  firm 
on  section,  and  there  was  marked  congestion  in  both,  while  it  was  more 
pronounced  in  one  kidney  than  the  other.  The  liver  was  congested,  the 
congestion  being  more  marked  in  patches. 

Sections  from  the  consolidated  upper  lobes  showed  under  the  micro- 
scope thickening  of  the  pleura  and  infiltration  with  round  cells.  The 
exudation  filled  the  alveoli,  and  was  breaking  down  in  some  cases  in  the 
centre.  The  vessels  were  injected,  and  there  were  haemorrhages  into  the 
alveoli.  The  periphery  of  the  lobules  was  infiltrated  with  round  cells. 
In  sections  of  the  kidney  there  was  slight  infiltration  around  glomeruli 
and  arterioles  with  round  cells  ;  the  epithelium  in  the  convoluted  tubules 
was  granular  and  disintegrating  ;  there  was  haemorrhage  in  the  straight 
tubules,  and  engorgement  of  vessels.  In  sections  of  liver  the  inter-  and 
intra-lobular  vessels  were  engorged  ;  there  were  interlobular  collections 
of  round  cells  displacing  the  liver  cells,  and  the  interlobular  connective 
tissue  was  infiltrated  with  round  cells  ;  the  liver  cells  were  granular  and 
cloudy. 

There  can  be  no  doubt  from  the  symptoms  and  post-mortein 
appearances  that  this  calf  had  been  suffering  from  septicaemia  .is 
the  result  of  introducing  the  septic  virus  and  crust  subcutaneously 
in  the  groin. 

The  two  Wiltshire  cows  were  killed,  and  there  was  nothing  of 
importance  to  note  in  one,  but  in  the  other  an  incision  into  the 
udder  revealed  an  enormous  abscess. 

18 


274  INFECTIVE   DISEASES. 

Though  the  naked-eye  appearances  of  the  kidney  in  this  case 
were  practically  healthy,  the  results  of  examining  sections  of  the 
kidney  under  the  microscope  were  extremely  instructive  and  interest- 
ing, as  they  showed  that  marked  changes  had  taken  place  which 
were  indicative  of  septic  complication. 

The  sections  showed  glomerulo- nephritis ;  there  was  infiltration 
of  the  capsule  of  Bowman  with  round  cells  ;  there  was  infiltration 
also  of  the  sheaths  of  the  vessels  with  round  cells,  especially  in 
the  cortex.  The  blood-vessels  in  the  boundary  zone  of  the  medulla 
were  engorged,  the  arterioles  of  the  glomeruli  were  also  engorged, 
and  there  were  slight  haemorrhages  into  the  capsule.  The  epithelium 
of  the  convoluted  tubules  was  granular,  opaque,  and  in  some  parts 
breaking  down. 

Sections  of  the  ulcers  of  the  teats  of  these  cows  were  also 
carefully  examined,  and  the  appearances  corresponded  exactly  with 
the  description  given  by  Klein. 

On  visiting  the  farms  it  was  found  that  there  were  altogether 
about  a  hundred  and  sixty  cows.  Only  a  few  had  proved  refractory, 
and  had  not  taken  the  disease  at  all.  The  rest  had  contracted  the 
disease  in  varying  degrees  of  severity.  About  fifty  at  a  time  were 
dry,  and  they  escaped  until  they  were  in  milk  again.  The  milk 
was  drunk  on  the  farms  and  in  the  village,  and  a  quantity  was 
supplied  to  a  large  town.  Most  careful  inquiries  were  instituted  to 
ascertain  the  existence  of  scarlatina  among  consumers  of  the  milk. 
So  far  the  research  was  completely  analogous  to  the  Hendon 
investigation;  but,  in  spite  of  the  contamination  of  the  milk,  no 
cases  of  scarlatina  were  found  either  on  the  farms  or  in  the  village, 
and  there  was  no  epidemic  in  the  town  in  which  the  milk  was 
distributed. 

The  disease,  in  fact,  was  cow-pox,  and  in  no  way  connected  with 
scarlet  fever ;  and  to  assist  others  who  may  undertake  a  similar 
inquiry  the  details  will  now  be  given  of  the  author's  investigation 
into  the  nature  of  the  outbreak  in  Wiltshire. 

THE  DISEASE  PROVED  TO  BE  COW-POX. 

Locality  of  the  Wiltshire  Outbreak. — There  is  considerable  interest 
attached  to  the  fact  that  the  farms  were  situated  a  few  miles  from 
Cricklade.  They  are  close  to  the  borders  of  Gloucestershire,  and  about 
twenty-five  miles  from  Berkeley.  They  are,  therefore,  within  that 
district  in  which  in  Jenner's  time  cow-pox  was  particularly  prevalent. 

Time  of  Year. — The  outbreak  commenced  about  the  end  of  September 
1886,  and  lasted  until  about  the  middle  of  December.  In  an  outbreak 


AN   OUTBREAK   OF   COW-POX.  275 

in  1885,  a  few  miles  from  these  farms,  but  on  a  separate  estate,  the 
disease  appeared  in  June  and  July. 

Origin  of  the  Outbreak. — The  author  made  careful  inquiries  as  to  the 
origin  of  the  outbreak,  but  beyond  ascertaining  with  certainty  that  the 
disease  appeared  first  at  one  farm,  and  was  conveyed  from  this  to  the 
other  farms,  all  evidence  was  negative.  The  milkers  were  unable  to 
say  whether  it  commenced  in  one  particular  cow  or  whether  it  broke 
out  in  several  simultaneously. 

The  only  information  which  could  be  obtained,  which  was  very 
suggestive,  was  that  the  milkers  were  in  the  habit  of  receiving  their 
friends  from  neighbouring  farms  on  Sundays.  The  friends  would  assist 
in  the  milking,  to  get  the  work  done  as  quickly  as  possible  on  these 
occasions.  As  it  was  reported  that  the  same  disease  had  occurred 
that  summer  on  a  neighbouring  farm,  it  is  quite  possible  that  it  was 
introduced  by  one  of  the  milkers'  friends. 

Mode  of  Dissemination. — When  the  disease  made  its  first  appearance, 
the  bailiff,  attributing  it  to  the  farm  being,  for  some  reason,  unhealthy, 
decided  to  remove  the  cows  to  other  farms.  The  herd  was  therefore 
divided  and  sent  to  two  other  farms.  From  these  cows  the  disease  was 
communicated  to  healthy  cows,  and,  as  this  interchange  was  repeated, 
not  only  of  the  cows,  but  of  the  milkers,  the  disease  was  communicated 
to  four  separate  farms. 

In  all  cases  the  disease  was  limited  to  the  teats,  and  was  conveyed 
from  the  teats  of  a  diseased  cow  to  the  teats  of  a  healthy  cow  by  the 
hand  of  the  milker.  In  no  case  was  there  any  evidence  of  the  disease 
being  produced  in  healthy  cows  by  other  means  than  contact. 

Bulls  and  dry  cows  remained  free  from  the  disease,  while  the  cows 
in  milk,  numbering  about  a  hundred  and  twenty,  were  all  attacked,  with 
the  exception  of  about  a  dozen,  which  proved  to  be  entirely  refractory. 

These  facts  explain  how  it  is  that  the  disease  has  been  known  from 
time  immemorial  as  the  "cow-pox."  We  never  hear  of  cattle-pox  or 
bull-pox.  We  have  not,  in  other  words,  to  deal  with  an  infectious 
disease  like  cattle-plague  or  pleuro-pneumonia,  but  with  a  disease  which 
is  communicated  solely  by  contact. 

The  disease  was  only  observed  in  the  cows  in  milk,  and  was  limited 
to  the  parts  which  come  in  contact  with  the  hand  of  the  milker.  The 
virus  was  mechanically  transferred  from  diseased  to  healthy  cows,  being 
communicated  to  all,  or  nearly  all,  the  animals  in  the  same  shed, 
whether  the  milker  had  vesicles  on  his  hand  or  not. 

Character  of  the  Erujrtion  on  the  Cow. — In  a  recent  case  which  was 
carefully  examined  the  teats  were  visibly  inflamed,  partly  red  and  partly 
livid  in  colour.  On  each  teat  there  were  vesicles,  some  broken,  and 
others  which  appeared  to  be  just  forming.  In  other  cases  there  was 
nothing  more  than  the  remains  of  broken  and  dried  vesicles,  and  more 
or  less  characteristic  crusts  on  the  teats. 

On  visiting  a  byre  at  the  time  that  the  cows  were  brought  in  to 
be  milked,  it  was  a  striking  sight  to  look  along  the  line  and  see  one 
animal  after  another  affected  with  the  eruption  ;  and  thus  one  character 


276  INFECTIVE   DISEASES. 

of  the  disease  was  clearly  shown — the  tendency  to  spread  through  a 
whole  herd. 

On  examining  the  eruption  carefully,  the  degree  of  severity  was 
found  to  differ  very  much  in  different  animals.  In  a  few  cases  the 
condition  was  most  distressing,  both  to  the  cow  and  to  the  observer.  In 
such  cases  the  teats  were  encrusted  with  huge,  dark  brown  or  black 
crusts,  which,  when  handled  in  milking,  were  broken  and  detached, 
exposing  a  bleeding,  suppurating,  ulcerated  base.  Such  ulcers  varied 
in  size  from  a  shilling  to  a  florin,  and  in  form  were  circular,  ovoid,  or 
irregular.  Weeks  afterwards,  when  the  animals  had  recovered,  the 
site  of  these  ulcers  was  marked  by  irregular  scars. 

All  the  milkers  agreed  as  to  the  general  characters  of  the  malady, 
laying  particular  stress  on  the  teats  being  red,  swollen,  and  painful  when 
handled.  Vesicles  would  then  appear  on  the  teats — two,  three,  four,  or 
more  on  each  teat.  They  were  soon  broken  in  milking,  and  irritated  into 
sores,  which  became  covered  with  thick  crusts.  From  four  to  six  weeks 
elapsed  before  they  had  entirely  healed.  Other  more  observant  milkers 
insisted  that  before  the  teats  were  red  and  swollen,  spots  or  pimples  first 
appeared  which  came  to  a  head.  This  head  increased  if  it  was  not  broken, 
which  might  be  the  case  if  it  was  situated  between  the  bases  of  the  teats, 
until  it  formed  a  greyish  vesicle  of  the  size  of  a  threepenny-piece  or 
even  larger. 

General  Symptoms  in  the  Cow. — As  to  the  general  condition  of  the  cows 
nothing  abnormal  was  observed.  They  appeared  in  the  best  of  health, 
and  in  only  one  particular  was  any  difference  from  their  condition  in 
health  stated  to  exist.  This  was,  that  in  the  majority  of  the  cases  there 
could  be  no  doubt  that  the  milk  had  diminished.  This  might  escape 
notice  by  inexperienced  milkers  in  any  particular  animal,  but  the  total 
amount  of  milk  supplied  by  the  herd  was  considerably  below  the  average. 

History  of  the  Eruption  communicated  to  the  Milkers. — The  most  striking 
characteristic  of  this  outbreak  was  the  communicability  of  the  disease 
to  the  milkers.  A  milker,  with  vesicles  which  presented  typically  the 
characters  of  casual  cow-pox,  was  taken  to  London  and  kept  under 
observation.  The  various  cases  will  be  described  in  the  order  in  which 
they  first  presented  themselves,  their  history  being  given  as  much  as 
possible  in  their  own  words. 

CASE  I. — J.  R.,  milker,  informed  the  author  that  he  was  the  first  to  catch 
the  eruption  from  the  cows.  He  stated  that  it  came  as  a  hard,  painful  spot, 
which  formed  "  matter  "  and  then  a  "  big  scab."  He  had  been  inoculated 
about  seven  weeks  previously.  He  pointed  to  the  scar  which  remained 
on  his  right  hand.  This  scar  presented  the  characters  of  an  irregular 
cicatrix,  indicating  considerable  loss  of  substance.  He  stated  that  he  had 
also  two  places  on  his  back,  where  he  supposes  he  had  inoculated  himself 
by  scratching.  He  had  continued  milking  ever  since,  but  had  had  no 
fresh  places. 

CASE  II.— W.  H.,  milker.  He  stated  that  he  was  inoculated  from  the 
cows  about  the  same  time  as  J.  R.  They  were  the  two  milkers  of 
the  herd  in  which  the  cow-pox  first  made  its  appearance.  The  eruption 


AN   OUTBREAK   OF   GOW-POX.  277 

appeared  in  one  place  on  each  hand.     He  pointed  to  two  irregular  scars 
as  the  remains  of  the  eruption. 

CASE  III.— J.  L.,  milker,  stated  that  he  also  caught  the  disease  from 
the  cows.  On  his  right  hand  a  spot  appeared  which  formed  a  blister, 
then  discharged  matter  and  produced  a  bad  sore.  Lumps  formed  at  the 
bend  of  his  elbow  and  in  his  armpit.  He  lost  his  appetite,  felt  very 
poorly,  and  was  obliged  to  leave  off  work  for  two  or  three  days. 

CASE  IV. — W.  K.,  a  labourer  on  the  farm,  was  put  on  as  a  milker  to 
take  the  place  of  one  of  the  others  with  bad  hands.  After  his  fifth  or 
sixth  milking — that  is  to  say,  about  three  days  after  first  milking  the  cows, 
—pimples  appeared  on  his  hands,  which  became  blistered  and  then  ran  on 
to  bad  sores.  He  pointed  to  three  irregular  scars  on  the  first  and  third 
fingers  and  palm  of  the  right  hand.  Lumps  appeared  in  his  elbow  and  in 
his  armpit,  but  he  did  not  feel  very  poorly  in  consequence. 

CASE  V.— J.  F.,  milker,  stated  that  about  a  month  ago  he  noticed 
spots  which  appeared  on  both  hands.  His  fingers  swelled  and  were  pain- 
ful. He  said  it  came  first  like  a  pimple,  and  felt  bard.  Then  it  "  weeped 
out"  water  in  four  or  five  days.  There  were  red  marks  creeping  up  to 
his  arm.  There  was  a  sort  of  throbbing  pain,  and  he  could  not  sleep  at 
night.  On  the  right  hand  there  was  a  scar,  but  on  the  left  hand  there 
was  an  ulcer  about  the  size  of  a  shilling  covered  with  a  thick  black  crust. 
The  crust  was  partially  detached,  and  exposed  a  granulating  ulcer.  It 
was  in  this  stage  the  exact  counterpart  of  the  ulcers  on  the  cow's  teats. 

CASE  VI.— W.  H.,  junior,  milker,  stated  that  he  had  both  hands  bad 
about  a  month  previously  :  first  on  the  index  finger  of  the  left  hand, 
and  then  on  the  right  hand  on  his  knuckle  and  between  the  first  and 
second  fingers.  He  said  that  it  came  up  like  a  hard  pimple,  and  the 
finger  became  swollen  and  red.  After  a  few  days  it  "  weeped  out " 
water,  and  then  matter  came  away.  Both  his  arms  were  swollen,  bat 
his  left  arm  was  the  worst.  About  a  fortnight  after,  he  noticed  kernels 
in  his  armpits,  which  were  painful  and  kept  him  awake  at  night.  His 
arms  became  worse,  he  could  not  raise  them,  and  he  had  to  give  up 
milking.  He  also  had  had  a  "  bad  place  "  on  the  lower  lip.  On  examina- 
tion, I  found  that  the  axillary  glands  were  still  enlarged  and  tender. 
He  volunteered  the  statement  that  the  places  were  just  like  the  sore 
teats. 

CASE  VII. — J.  H.,  the  bailiff's  son,  also  milked  the  cows.  He  had 
a  sore  on  the  upper  lid  of  his  right  eye  and  on  his  left  hand.  In  both 
cases  he  had  been  previously  scratched  by  a  cat,  and  the  scratches  were 
inoculated  from  the  cow's  teats.  The  right  hand  also  had  been  inoculated. 
The  eruption  broke  out  a  fortnight  previously.  His  hands  were  swollen, 
red,  and  hot.  He  felt  very  poorly  and  went  to  bed.  Little  spots  like 
white  blisters  appeared  on  the  back  of  his  right  hand.  His  mother 
remarked  that  they  "rose  up  exactly  as  in  vaccination."  Thick  dark 
brown  scabs  formed.  He  was  very  ill  for  two  or  three  days,  but  did  not 
send  for  a  doctor.  He  had  painful  lumps  at  the  bend  of  his  arm  and  in 
the  armpit.  He  gave  up  milking,  and  had  not  taken  to  it  since. 

On  examining  him,  the  thick  crusts  on  his  right  hand  were  identical 


(UNIVERSITY 

^^r^  Pir  irm 


278  INFECTIVE   DISEASES. 

with  the  stage  of  scabbing  in  vaccinia.  The  scabs  fell  off  in  about  three 
weeks  to  a  month,  and  left  permanent,  depressed  scars. 

CASE  VIII. — W.  P.,  milker.  This  case  was  pointed  out  on  the  occasion 
of  another  visit,  and  is  the  only  one  in  which  the  eruption  was  seen  in 
its  earlier  stages. 

The  history  of  this  boy  is  as  follows.  He  had  taken  the  place  of  one 
of  the  other  milkers  who  had  vesicles  on  his  fingers,  and  had  been  obliged 
to  give  up  milking.  After  the  seventh  time  of  milking  he  noticed  a 
small  pimple  on  his  right  cheek.  This  became  larger  and  vesicular. 
On  examination  it  presented  a  depressed  vesicle  with  a  small  central 
yellowish  crust  and  a  tumid  margin,  the  whole  being  surrounded  by  a 
well-marked  areola  and  considerable  surrounding  induration.  On  raising 
the  central  incrustation  a  crater-like  excavation  was  seen,  in  which 
lymph  welled  up  and  trickled  down  the  boy's  cheek.  On  the  following 
day  the  crust  had  re-formed,  and  was  studded  with  coagulated  lymph. 
The  areola  became  more  marked,  and  on  pricking  the  margin  of  the 
vesicle,  the  exuding  contents  were  slightly  turbid. 

From  this  day  the  surrounding  infiltration  increased  enormously, 
the  whole  cheek  was  inflamed,  and  the  eyelids  so  oedematous  that  the 
eye  was  almost  closed.  There  was  enlargement  of  the  neighbouring 
lymphatic  glands.  The  crust  which  had  re-formed  thickened  day  by  day. 
It  retained  the  character  of  central  depression,  and  was  situated  on  a 
reddened,  raised,  and  indurated  base  (Plate  VII.). 

From  this  date  the  surrounding  induration  gradually  diminished. 
The  crust  changed  in  colour  from  dark  brown  to  black,  and  finally 
fell  off,  leaving  an  irregular,  depressed  scar.  This  scar,  when  seen  several 
months  afterwards,  was  found  to  be  a  permanent  disfigurement.  The 
eruption  appeared  on  the  fourth  day  after  exposure  to  infection,  and 
allowing  two  days  for  incubation,  the  vesicle  was  at  its  height  on  the 
seventh  or  eighth  day,  and  a  typical  tamarind- stone  crust  fell  off  on 
the  twenty-first  day  after  infection,  leaving  a  depressed,  irregular 
cicatrix. 

A  vesicle  also  formed  on  the  thumb  of  the  left  hand.  Two  days 
after  the  pimple  appeared  on  his  cheek,  the  lad  said  that  he  first  noticed 
a  pimple  on  his  thumb,  and  this,  on  examination,  presented  a  greyish 
flattened  vesicle,  about  the  size  of  a  sixpence.  Later,  its  vesicular 
character  was  much  more  marked,  and  a  little  central  crust  had  com- 
menced to  form.  The  margins  became  very  tumid,  giving  it  a  marked 
appearance  of  central  depression.  The  vesicle  was  punctured  at  its 
margin  with  a  clean  needle,  and  from  the  beads  of  lymph  which  exuded 
a  number  of  capillary  tubes  were  filled. 

Two  days  afterwards  suppuration  had  commenced,  the  vesicle  con- 
tained a  turbid  fluid,  and  the  areola  was  well  marked.  Later,  the  crust 
had  assumed  a  peculiar  slate- coloured  hue,  and,  on  pressing  it,  pus 
welled  up  through  a  central  fissure.  The  areola  had  increased,  and 
there  was  considerable  inflammatory  thickening.  The  lymphatic  glands 
in  the  armpit  were  enlarged  and  painful.  Though  there  was  deep 
ulceration,  which  left  a  permanent  scar,  the  ulceration  did  not  assume 


DESCRIPTION    OF    PLATE    VII. 
Casual  Cow-pox. 

FIG.  1. — Case  of  W.  P ,  a  milker,  infected  from  the  teats  of  a  cow  with 

natural  cow-pox.  There  was  a  large  depressed  vesicle  with  a  small 
central  crust  and  a  tumid  margin,  the  whole  being  surrounded  by  a 
well-marked  areola  and  considerable  surrounding  induration. 

FIG.  2. — The  same  case  a  week  later,  showing  a  reddish-brown  crust  on  a 
reddened  elevated  and  indurated  base. 


Plate  TOT. 


Eg  2. 


CASUAL     COW-POX. 


OF  THB 

UNIVERSITY 

OF  r 


AN    OUTBREAK   OF   COW-POX.  279 

quite  so  severe  a  character  as  in  some  of  the  other  milkers.  Possibly 
this  may  be  accounted  for  to  some  extent  by  the  fact  that  the  pock  was 
covered  with  a  simple  dressing  instead  of  being  subjected  to  the  irritation 
and  injury  incidental  to  working  on  the  farm. 

Re-vaccination  of  the  Milker*. — There  were  in  all  eight  milkers,  varying 
in  age  from  seventeen  to  fifty-five,  who  had  vesicles  on  their  hands 
from  milking  the  cows.  Seven  had  been  vaccinated  in  infancy,  but  not 
since  ;  one  had  been  revaccinated  on  entering  the  navy  at  fifteen.  They 
were  all  revaccinated  by  a  public  vaccinator  after  complete  recovery 
from  the  casual  cow-pox  (that  is  to  say,  from  three  to  four  months 
afterwards),  and  were  all  completely  protected.  On  the  other  hand, 
two  of  the  three  milkers  who  had  escaped  infection  from  the  casual 
cow-pox  were  also  vaccinated,  with  the  result  in  one  of  typical 
revaccination,  in  the  other  of  very  considerable  local  irritation. 

Retro-vaccination  of  Calves. — The  result  of  retro-vaccinating  calves  with 
the  humanised  lymph  was  strictly  in  accordance  with  the  experience  of 
Ceely,  who  has  pointed  out  that  in  retro-vaccination  from  the  milker's 
hands  the  results  are  doubtful,  and  depend  greatly  on  the  animals  selected. 
"Those  of  a  light  colour  and  with  thin  skins  were  generally  preferred, 
but  often  without  avail,  scarcely  one-half  of  the  operations  succeeding." 
"  Vaccine  lymph,  in  passing  from  the  cow  to  man,  undergoes  a  change 
which  renders  it  less  acceptable  and  less  energetic  on  being  returned  to 
many  individuals  of  the  class  producing  it  ;  some  refuse  it  altogether." 
Two  cases  out  of  four  succeeded,  and  an  eruption  was  produced  with 
all  the  typical  characters  of  vaccinia,  but  running  rather  a  rapid  course, 
and  the  protection  passing  off  after  a  few  weeks,  while  the  result 
obtained  in  calves  inoculated  with  pus  or  scrapers  from  ulcers  was  in 
accordance  with  what  is  well  known  to  occur  if  pus  instead  of  lymph 
is  taken  for  carrying  on  calf  to  calf  vaccination. 

That  the  cow-pox  in  Wiltshire  was  identical  with  the  so-called 
Hendon  cow-disease  there  can  be  little  room  for  doubt,  for  in  both 
cases  we  find  that — 

1.  The  disease  spread  through  a  whole  herd  of  milch  cows. 

2.  The  disease  was  characterised  by  the  appearance  of  vesicles, 
which   were  broken  by   the  hand  of  the  milker,  and  irritated  into 
deep  ulcerations. 

3.  The  disease  was  conveyed  from  one  cow  to  another  by   the 
hand  of  the  milker. 

4.  The  vesicular  eruption   was   communicable   to   the   hand  of 
the  milker. 

5.  The  disease  was  not  fatal,  and  in  cows  which  were  killed  and 
examined  the  post-mortem  appearances  could  not  be  distinguished 
from  accidental  complications. 

6.  The  naked-eye  appearances  and  the  duration  of  the  ulcers  of 
the  teats  were  the  same. 


280  INFECTIVE   DISEASES. 

7.  Sections  of  the  ulcers  showed  under  the  microscope  identical 
appearances  of  a  cellular  character,  and  the  purulent  discharge  of 
the  ulcers  contained  pyogenic  cocci. 

8.  The  results  produced  by  inoculation  of  calves  with  the  septic 
virus  were  identical. 

If  we  examine  the  chain  of  argument  which  has  been  brought 
forward  to  maintain  the  existence  of  cow-scarlatina  at  Hendon,  we 
find  that  it  was  urged  : — 

1.  That  the  Hendon  cow  disease    was  a  disease   in  which   the 
post-mortem  appearances  resembled  scarlatina. 

2.  That  this  disease  was  associated  with  a  streptococcus,  which 
produced,    by   inoculation    in    calves,    a   disease    with    post-mortem 
appearances  similar  to  those  of  the  Hendon  cows. 

3.  That   a   streptococcus    regarded   as   identical    with    the   one 
above  mentioned  was  found  in  certain  cases  of  scarlatina  in  man, 
which  when  inoculated  in  calves  produced  post-mortem  appearances 
similar  to  the  post-mortem  appearances  in  the  original  Hendon  cows 
and  in  certain  cases  of  scarlatina  in  man. 

But  the  microscopical  appearances  of  the  kidney  of  a  Wiltshire 
cow  were  identical  with  those  which  were  regarded  as  indicating 
scarlatina  in  a  Hendon  cow ;  and,  indeed,  the  statements  as  to  the 
post-mortem  appearances  in  the  Hendon  cows,  when  studied,  not 
only  do  not  necessarily  indicate  scarlatina,  but  they  cannot  even  be 
considered  of  primary  importance,  or  as  throwing  much  light  on  the 
question  of  scarlatina  at  all.  The  description  of  the  naked- eye 
appearances  in  both  cows  only  suggests  coincident  pleurisy  or 
pleurisy  with  pneumonia.  The  microscopical  appearances  in  both 
were  suggestive  of  septic  complication. 

A  careful  examination  of  the  post-mortem  appearances  of  calves 
inoculated  with  scraping  of  an  ulcer  of  a  Hendon  cow,  or  with 
cultivations  of  the  streptococcus  from  certain  cases  of  scarlatina, 
brings  to  light  much  more  striking  changes.  These  appearances, 
however,  cannot  be  regarded  as  indicative  of  scarlatina.  They  are 
in  reality  the  post-mortem  appearances  of  septic  poisoning,  and 
occur  commonly  in  many  diseases.  This  is  clearly  shown  by  com- 
paring the  post-n.ortem  appearances  in  the  calf  which  was  killed 
while  suffering  from  septicaemia  as  the  result  of  inoculation  from 
the  ulcers  of  a  Wiltshire  cow.  These  visceral  changes  are  not  to 
be  distinguished  from  the  post-mortem  appearances  described  in 
the  calves  inoculated  by  Klein,  Consequently,  that  the  strepto- 


AN   OUTBREAK   OF   COW-POX.  281 

coccus  found  in  certain  cases  of  scarlet  fever  should  produce  on 
inoculation  in  calves  certain  post-mortem  appearances  which  are 
found  in  many  diseases,  and  should  fail  to  produce  fever,  ulceration 
of  the  tonsils,  or  scarlatinal  rash,  or  any  condition  in  the  least 
resembling,  clinically,  the  disease  in  man,  and  yet  that  the  result 
should  be  regarded  as  scarlatina  in  the  calf,  is  a  conclusion  quite 
untenable. 

It  is  true  that  visceral  lesions  similar  in  character  were  produced 
in  calves  whether  inoculated  with  scrapings  or  with  streptococci  from 
ulcers  of  the  Hendon  cows  or  with  streptococci  from  certain  cases  of 
scarlet  fever.  In  both  cases  the  streptococcus  is  pathogenic,  and 
inoculation  of  Streptococcus  pyogenes  or  the  inoculation  of  septic 
virus,  is  liable  to  produce  septicaemia,  These  facts  constitute  a  mass 
of  evidence  which  justifies  the  conviction  that  the  pathological  data 
which  appeared  to  support  the  theory  that  the  vesicular  disease  of 
the  teats  of  cows  at  Hendon  was  scarlatina  in  the  cow,  admit  of  an 
entirely  different  interpretation^  and  there  can  be  no  longer  any 
doubt  that  the  milk  was  not  infected  by  the  cows  but  with  the  virus 
of  scarlet  fever  from  some  human  source  which  Mr.  Power  failed  to 
discover. 

All  the  other  evidence  reported  to  the  Board  of  Agriculture  pointed 
to  the  same  conclusion.  The  disease  at  Hendon  was  admittedly 
introduced  from  Derbyshire ;  and  from  Professor  Axe's  report  it 
appears  that  only  a  part  of  the  herd  was  sold  to  the  farmer  at 
Hendon;  other  cows  with  the  same  eruption  were  transferred  to 
other  dairy  farms,  and  the  disease  communicated  to  healthy  cows  as 
at  Hendon,  but  in  no  instance  did  scarlet  fever  occur  among  the 
consumers  of  the  milk.  At  the  farm  of  the  brother  of  the  dealer  the 
disease  was  communicated  to  three  of  the  milkers,  and  the  eruption 
diagnosed  by  Dr.  Bates  as  vaccinia. 

All  this  evidence  must  be  regarded  as  conclusive.  The  con- 
tamination of  the  milk  at  Hendon  with  scarlet  fever  must  neces- 
sarily have  been  a  mere  coincidence ;  and  the  conclusion  that 
the  milk  could  not  possibly  have  become  infected  from  any 
human  source  is  untenable.  Professor  Axe  even  ascertained 
that  scarlet  fever  existed  at  Hendon  during  several  months  of 
1885,  and  that  the  dwellings  where  cases  occurred  stood  within  MX 
hundred  yards  of  the  cowsheds  which  contained  the  incriminated 
cows,  and  that  out  of  fourteen  men  on  the  farm  six  lived  in 
a  district  where  cases  occurred.  Professor  Axe  has  also  stated 
that  the  father  and  brother  of  a  girl  with  scarlet  fever,  visited  the 
dairy  during  her  illness.  Whether  any  of  those  engaged  on  the 


282  INFECTIVE   DISEASES. 

farm  suffered  from  latent  scarlet  fever  does  not  appear  to  have 
been  ascertained. 

There  is,  it  is  true,  no  evidence  to  show  that  any  one  daily  carried 
infection  to  the  milk,  but  the  exact  path  of  infection  is  not  always 
easy  to  trace  ;  and  because  it  was  not  actually  traced  it  was  hardly 
reasonable  to  assume  that  the  possibility  of  contamination  from  a 
human  source  could  be  altogether  eliminated. 

In  attempting  to  communicate  scarlet  fever  to  cows  Professor 
M'Fadyean  confirmed  the  negative  results  which  had  been  experi- 
enced in  some  earlier  experiments  by  Klein.  In  1882  Klein 
inoculated  and  fed  cows  and  yearling  heifers  with  diseased  products 
from  human  patients,  using  desquamated  cuticle  and  the  discharges 
from  the  throat ;  but  the  experiments  all  failed.  M'Fadyean's 
failures  were  still  more  marked.  Cows  and  calves  were  inoculated 
with  blood  from  scarlet  fever  patients,  and  they  were  made  to  drink 
water  thickened  with  desquamated  cuticle,  but  all  the  experiments 
proved  unsuccessful. 

The  author  believes  that  the  outbreak  at  Hendoii  was  one 
of  cow-pox,  which  was  prevalent  in  this  country  in  1886.  The 
outbreak  in  Wiltshire  could  not  be  distinguished  bacteriologically 
or  clinically  or  in  its  micropathology,  from  the  disease  at  Hendon, 
and  the  Wiltshire  outbreak  proved  on  investigation  to  be  true  cow- 
pox.  This  conclusion  was  questioned  at  the  time,  as  cow-pox  was 
generally  believed  to  be  extinct  in  England  ;  but  that  view  is 
entirely  fallacious,  and  the  author's  conclusions  have  since  been  fully 
confirmed  by  independent  observers,  whose  work  will  be  referred  to 
in  another  chapter  (p.  321). 

Stamping-out  System. — The  Notification  Act  of  1890  may  be 
voluntarily  adopted  in  sanitary  districts,  but  it  would  be  a  great 
advantage  if  notification  were  carried  out  uniformly  all  over  the 
country.  Prompt  information  may  lead  to  detecting  the  origin  of 
cases  of  scarlet  fever,  and  isolation  and  disinfection  will  assist  in  pre- 
venting its  spread.  Epidemics  have  occurred  on  a  large  scale  owing 
to  scarlet  fever  existing  among  those  engaged  in  dairy  work,  and 
the  precaution  not  being  taken  of  stopping  the  milk  supplied  to  the 
consumers.  Scarlet  fever  cannot  be  so  readily  controlled  as  small- 
pox, for  it  may  be  spread  by  mild  cases  before  the  nature  of  the 
disease  is  suspected,  ancl  small-pox  cannot  be  conveyed  in  milk. 


MEASLES.  283 

MEASLES. 

Measles  is  a  contagious  disease  peculiar  to  man.  It  lasts  for 
one  or  two  weeks,  and  produces  fever,  catarrh  of  the  respiratory 
mucous  membrane,  and  a  characteristic  rash.  It  is  highly  contagious, 
especially  before  the  nature  of  the  disease  is  revealed;  there  is 
consequently  great  difficulty  in  preventing  its  spread  in  schools 
and  households.  The  contagium  appears  to  be  given  off  from  the 
body,  principally  if  not  entirely,  by  the  breath.  One  attack  is  pro- 
tective against  future  attacks.  The  whole  population  of  a  country 
may  acquire  a  certain  degree  of  immunity.  Measles  introduced  into 
countries  where  it  was  previously  unknown  assumes  a  most  malig- 
nant form.  There  are  no  characteristic  post-mortem  appearances. 

Bacteria  in  Measles. — Micrococci  have  been  found  in  the 
blood,  catarrhal  exudation,  and  skin,  by  Keating,  Babes,  and  others, 
but  they  are  accidental  epiphytes  of  no  importance,  or  associated 
with  secondary  complications,  as  in  scarlet  fever. 

Canon  and  Pielicke  have  found  in  the  blood  small  bacilli  varying 
in  form.  They  do  not  grow  on  nutrient  agar  or  blood  serum,  but 
cultures  were  obtained  by  pricking  the  finger  of  a  patient  suffering 
from  measles,  and  allowing  the  blood  to  drop  into  sterilised  broth. 
After  a  few  days  the  broth  became  cloudy,  and  later,  a  flocculent 
deposit  formed.  The  bacilli  were  also  obtained  from  the  nasal  and 
conjunctival  secretions.  The  nature  of  the  contagium  of  measles 
is  unknown. 

Stamping- out  System. — Measles  is  not  easily  controlled  by  the 
stamping-out  system ;  it  is,  in  fact,  extremely  difficult,  almost  impos- 
sible, to  prevent  its  spread,  as  it  is  especially  infectious  during  the 
period  of  incubation.  Notification,  isolation,  and  disinfection  assist  in 
controlling  an  epidemic,  but  the  value  of  the  system  does  not  apply 
to  the  same  extent  in  measles  as  in  other  infectious  diseases. 


CHAPTER   XX. 

SMALL-POX. — CATTLE   PLAGUE. 

SMALL-POX. 

SMALL-POX  is  an  infections  and  inoculable  disease  of  man,  charac- 
terised by  sadden  and  severe  fever,  followed  in  forty-eight  hours  by  a 
characteristic  papular  eruption  which  gradually  becomes  vesicular 
and  then  pustular.  The  virus  is  contained  in  the  vesicles,  and  in  a 
concentrated  form  in  mature  pustules.  It  also  passes  into  the  air 
from  the  breath  and  skin.  Infection  may  occur  from  the  dead  body, 
and  clothes  and  bedding  may  retain  the  contagium  for  months. 
One  attack,  as  a  rule,  gives  immunity  against  future  attacks. 

Small-pox  is  undoubtedly  a  disease  foreign  to  this  country.  Its 
home  is  in  the  East.  Some  of  the  old  writers  held  that  it  spread  to 
Europe  from  Alexandria  about  the  year  640  A.D.,  following  in  the 
wake  of  the  Arab  conquests  in  Egypt,  Palestine,  Persia,  along  the 
Asiatic  coast,  through  Lycia,  Gallicia,  along  the  coast  of  Africa, 
and  across  the  Mediterranean  to  Spain  ;  others  maintained  that  it 
was  not  introduced  until  the  end  of  the  eleventh  or  beginning  of  the 
twelfth  century,  by  the  returning  Crusaders.  At  any  rate,  small- 
pox was  imported  from  the  East,  and  probably  from  Egypt.  Hero- 
dotus, who  visited  Egypt,  leads  us  to  infer  that  epidemics  were 
unknown  there  during  the  rule  of  the  Pharaohs ;  but  Egypt 
undoubtedly  became  a  hotbed  of  pestilence  during  the  Mohammedan 
occupation.  Prosper  Alpinus  imagined  that  both  the  plague  and  the 
small-pox  were  concocted  in  the  putrid  waters  of  the  Nile,  but  he 
would  probably  have  been  more  correct  if  he  had  suggested  that 
they  arose  from  the  insanitary  condition  of  the  Arab  conquerors 
and  their  filthy  camp  followers,  who  did  their  best  to  destroy  all 
that  remained  of  that  magnificent  civilisation  which  had  existed  in 
the  days  of  the  ancient  Egyptians. 

We  do  not  know  the  exact  period  at  which  small-pox  was  first 
imported  into  England,  and  the  records  of  the  disease  are  very 
meagre  until  the  sixteenth  century. 

284 


SMALL-POX.  285 

In  1593  Simon  Kell \vaye  appended  to  his  work  on  the  Plague 
a  short  treatise  on  the  small-pox.  "  Oftentimes,"  he  wrote,  "  those 
that  are  infected  with  the  plague  are  in  the  end  -of  the  disease 
sometimes  troubled  with  the  small  pocks  or  measels,  as  also  by  good 
observation  it  hath  been  seen  that  they  are  fore-runners  or  warnings  of 
the  plague  to  come."  According  to  Kellwaye  the  disease  arose  from 
the  "  excrements  of  all  the  foul  humours  in  our  bodies,  which  striving 
with  the  purest  doth  cause  a  supernatural  heat  and  ebullition  of  our 
blood,  always  beginning  with  a  feaver  in  the  most  part." 

Small-pox  steadily  increased  in  the  seventeenth  century  until  it  was 
a  formidable  scourge,  for  no  advantage  was  taken  of  all  the  experi- 
ence which  had  been  gained  in  dealing  with  the  plague.  No  public 
measures  were  adopted  to  cope  with  the  disease,  and  the  people  came 
to  regard  the  new  pestilence  as  a  visitation  which  was  unavoidable. 
Early  in  the  eighteenth  century,  small-pox  inoculation  was  introduced, 
and  this  was  superseded  in  the  nineteenth  century  by  vaccination. 

Examination  of  small -pox  cases  after  death  does  not  reveal  any 
characteristic  lesions  in  the  internal  organs,  but  sections  of  small- 
pox vesicles  show  an  important  structure.  A  vesicle  is  formed  by 
the  exudation  raising  up  the  outer  layer  of  epidermis,  and  the  chief 
feature  is  the  formation  of  a  vacuolated  structure  in  which,  especially 
in  the  later  stages,  bacteria  are  found  in  abundance. 

Bacteria  in  Small-pox. — Cohn  and  Weigert  found  cocci  in 
variolous  lymph.  Hlava  found  Streptococcus  pyogenes  in  the  pustules, 
and  Garre  streptococci  in  the  internal  organs  in  a  case  of  variola 
hsemorrhagica.  In  a  fatal  case  of  variola  complicated  with  pemphigus 
Garre  found  a  streptococcus  in  the  pemphigus  vesicles.  Klein 
and  Copeman  have  found  a  small  bacillus  which  they  regard  as 
characteristic,  but  its  biological  characters  are  unknown,  as  it  will 
not  grow  on  any  nutrient  media.  The  bacteria  commonly  found  in 
variolous  pus  are  the  usual  pyogenic  organisms.  The  nature  of 
the  contagium  of  small-pox  is  unknown. 

Protective  Inoculation. — Experience  had  taught  that  a  person 
was  not,  as  a  rule,  attacked  with  small-pox  a  second  time;  but  when 
and  how  the  method  of  artificially  inducing  a  mild  form  -of  the 
disease  was  discovered,  or  when  this  preventive  treatment  was  first 
employed,  is  unknown.  Avicenna  of  Bokhara  was  credited  with  the 
discovery,  and  it  was  supposed  that  the  practice  was  carried  by 
Tartar  and  Chinese  traders  to  Surat,  Bengal,  and  China,  and  by 
the  Mahommedan  pilgrims  to  Mecca.  In  Constantinople  it  was 
supposed  by  some  to  have  been  introduced  from  the  Morea  by 
an  old  woman,  and  by  others  by  the  women  of  Circassia.  The 


286  INFECTIVE   DISEASES. 

Circassian  women  fastened  three  needles  together,  and  pricked  the 
skin  over  the  pit  of  the  stomach  and  heart,  the  navel,  the  right 
wrist,  and  the  left  ankle.  The  variolous  matter  was  applied  to  the 
bleeding  points,  and  the  eruption  came  out  in  five  or  six  days.  In 
Constantinople  scarifications  were  made  on  the  forehead,  wrists, 
and  legs,  and  carefully  selected  virus  applied  to  the  incisions.  The 
needle  used  was  a  three-edged  surgeon's  needle,  or  the  operation  was 
performed  with  a  lancet.  The  virus  was  obtained  by  pricking  the 
vesicles,  and  pressing  out  the  matter  into  a  clean  glass  vessel.  The 
Armenians  preferred  to  be  inoculated  in  both  thighs.  In  Barbary 
a  slight  wound  was  made  between  the  thumb  and  forefinger,  and 
the  virus  obtained  from  a  mild  form  of  small-pox.  In  Hindustan 
the  operation  was  performed  at  certain  seasons  of  the  year,  and  a 
preparatory  regimen  enforced.  The  inoculators  were  very  careful 
in  the  selection  of  the  virus,  as  they  had  learnt  its  varying  intensity, 
and  they  were  credited  with  being  able  to  control  the  amount  of 
the  eruption.  They  preferred  to  inoculate  the  outside  of  the  arm, 
midway  between  the  wrist  and  the  elbow  in  males,  and  between  the 
elbow  and  the  shoulder  in  females.  The  skin  over  the  part  to  be 
inoculated  was  first  well  rubbed  with  a  piece  of  cloth  ;  then,  with 
slight  touches  of  a  small  instrument,  little  wounds  were  made  over 
an  area  which  might  be  covered  by  a  small  coin,  and  sufficient  to 
cause  just  an  appearance  of  blood.  A  pledget  of  cotton -wool 
charged  with  the  variolous  matter,  and  moistened  with  water,  was 
applied  to  the  wound.  This  virus  was  obtained  from  inoculated 
pustules  of  the  preceding  year. 

In  China  the  contents  of  the  variolous  pustules  were  dried  and 
kept  for  several  years.  If  the  virus  was  to  be  used  from  fresh 
pustules  the  "  acrimony  "  of  the  matter  was  corrected  by  steaming. 
The  dried  powder  was  made  into  a  paste,  which  was  wrapped  up  in 
cotton -wool  and  introduced  into  the  nostril. 

The  Greeks  were  more  cautious  in  their  procedure,  and  were 
said  to  inoculate  tens  of  thousands  without  an  accident.  They 
operated  only  upon  those  in  perfect  health,  punctures  were  made 
with  needles,  and  the  virus  was  used  in  the  crude  state,  freshly 
obtained  from  the  "  kindly  "  pustules  of  a  young  child.  They  were 
particularly  careful  in  the  choice  of  the  "ferment." 

Dr.  Perrot  Williams,  in  1722,  wrote  that  the  practice  of  com- 
municating small-pox  had  long  been  employed  in  South  Wales.  The 
oldest  inhabitants  said  that  it  had  been  a  common  practice  with 
them  "time  out  of  mind,"  but  Lady  Mary  Wortley  Montagu  was 
responsible  for  the  general  adoption  of  small-pox  inoculation  in 


SMALL-POX.  287 

England  by  persuading  physicians  in  London  to  employ  it.  Lady 
Mary  had  her  child  inoculated  in  Turkey.  An  old  Greek  woman  inocu- 
lated one  arm,  and  Mr.  Maitland,  surgeon  to  the  Embassy,  the  other. 
The  disease  ensued  in  due  course  with  an  eruption  of  a  hundred 
pustules.  This  was  the  first  time  that  the  Byzantine  method  of 
inoculation  was  performed  upon  an  English  subject.  In  1721  Dr. 
Harris  delivered  a  lecture  before  the  College  of  Physicians,  and 
described  the  successful  inoculation  of  four  children  of  the  French 
consul  at  Aleppo,  by  means  of  a  thread  imbued  with  variolous  pus. 
A  "daughter  of  Lady  Mary  was  inoculated  in  England  by  Maitland  in 
1721,  and  subsequently  a  number  of  criminals  were  inoculated  by 
him.  Incisions  were  made  through  the  cutis,  and  pledgets  which  had 
been  steeped  in  variolous  pus  from  ripe  pustules,  were  applied  to  the 
wound.  This  was  known  as  Maitland's  or  the  reformed  operation, 
but  it  was  soon  modified,  as  troublesome  ulcers  resulted.  Shortly 
afterwards  Maitland  encountered  another  obstacle.  The  child  of  a 
Mr.  Batt  was  inoculated,  had  plenty  of  pustules,  and  soon  recovered, 
but  six  of  Mr.  Batt's  domestic  servants,  "  who  all  in  turn  were 
wont  to  hug  this  child  while  under  this  operation,  and  whilst  the 
pustules  were  out,  never  suspecting  them  to  be  infectious,  were  all 
seized  at  once  with  the  right  natural  small-pox  of  several  and  very 
different  kinds." 

Dr.  Jurin  in  1729  reverted  to  the'  Eastern  method,  and  recom- 
mended virus  from  a  mild  case  of  small-pox,  but  the  virus  was  still 
taken  from  perfectly  maturated  pustules,  and  the  operation  continued 
to  be  followed  by  bad  results.  In  order  to  diminish  the  risks,  Burgess 
in  1766  advocated  certain  improvements.  An  incision  about  an  inch 
long  was  made  on  each  arm  through  the  cuticle,  but  not  so  deep  as 
to  wound  the  cellular  tissue.  A  variolous  thread  was  laid  along  the 
whole  length  of  the  wound  and  fixed  with  plaster.  Ulcerations  and 
other  accidents  continued  to  take  place,  and  a  new  epoch  in  the 
history  of  inoculation  was  the  introduction  of  the  Suttonian  method, 
in  1764-6. 

It  was  said  that  Mr.  Button,  with  his  assistants,  inoculated 
one  hundred  thousand  persons.  The  method  was  kept  secret  at  first, 
but  the  essential  points  were  all  discovered  and  published  by  Dr. 
Dimsdale.  Dimsdale  recommended  a  very  slight  puncture  with  a 
lancet  wet  with  variolous  matter.  Subsequently,  Sutton  published 
an  account  of  his  method,  and  the  result  of  his  operation  may  be 
given  in  his  own  words. 

"The  lancet  being  charged  with  the  smallest  perceivable  quantity 
(and  the  smaller  the  better)  of  unripe,  crude,  or  watery  matter,  immediately 


288  INFECTIVE    DISEASES. 

introduce  it  by  puncture,  obliquely,  between  the  scarf  and  true  skin, 
barely  sufficient  to  draw  blood,  and  not  deeper  than  the  sixteenth  part  of 
an  inch.  Neither  patting,  nor  daubing  of  the  matter,  in  or  over  the 
punctured  part,  is  at  all  necessary  to  its  efficacy.  This  practice,  indeed, 
is  rather  prejudicial  than  otherwise,  as  it  may  affect  the  form  of  the 
incision,  and  thus  be  apt  to  confound  our  judgment  upon  it. 

"  Indications  of  the  Incision. — In  the  incipient  state  of  variolous 
increase  in  the  incision,  a  small  florid  spot  appears  on  the  part  of  access, 
resembling  a  flea-bite  in  size  ;  and  on  passing  the  finger  lightly  over  it  a 
hardness  is  felt  not  larger  than  a  small  pin's  head.  This  florid  appearance 
and  hardness  denote  that  the  variolous  principle  is  effectually  imbibed, 
and  their  indications  point  no  farther,  unless  the  progress  to  vesication 
be  very  slow,  in  which  case  an  uncomfortable  number  of  pustules  may  be 
expected  to  follow.  The  florid  spot  in  most  instances  of  inoculation  is 
somewhat  larger,  or  more  extended  on  the  second,  than  on  the  third  day 
after  the  insertion. 

"About  the  fourth  day  from  inoculation,  should  the  incision  begin 
to  vesicate,  an  itching  sensation  will  be  complained  of  on  the  place  of 
insertion — the  occurrence  of  which  symptom  is  the  first  indication  of  a 
favourable  event,  yet  not  of  sufficient  importance  to  justify  any  present 
relaxation  in  the  preparatory  proceedings. 

"  The  vesication  of  the  incision  in  most  instances  will  begin  to  be  visible 
on  the  fourth  or  fifth  day  after  the  insertion  of  the  matter  ;  the  sooner 
it  becomes  so,  the  more  favourable  may  be  expected  to  be  the  event.  The 
extent  or  diameter  of  the  vesication  at  this  stage  does  not  usually  exceed 
that  of  a  large  pin's  head,  and  it  has  invariably  a  dint  or  small  depression." 

Adams  obtained  still  more  striking  results  by  inoculating  with 
variolous  lymph  from  pearl-pox,  a  mild  variety  of  small-pox.  Starting 
with  lymph  obtained  from  this  benign  form  of  small-pox,  and 
selecting  the  cases,  and  carrying  on  arm  to  arm  variolation,  the 
results  obtained  were  practically  identical  with  the  phenomena 
obtained  by  inoculation  of  the  arm  with  cow-pox  lymph.  Similar 
results  were  obtained  by  Guillou,  but  more  rapidly.  In  1827  there 
was  an  epidemic  of  variola,  and  Guillou,  having  no  vaccine  virus, 
took  variolous  lymph  from  a  girl  fifteen  years  of  age  on  the  fifth  clay 
of  the  eruption.  The  case  was  one  of  varioloid  or  mild  small-pox,  attri- 
buted to  previous  vaccination.  The  variolous  lymph  was  inserted  in 
ten  places  on  the  arm  of  a  healthy  infant  still  at  the  breast.  This 
inoculation  produced  ten  beautiful  "  vaccine  "  vesicles,  from  which,  on 
the  ninth  day,  forty -two  infants  were  inoculated  under  the  eyes  of 
two  of  the  local  authorities.  These  furnished  virus  for  the  inoculation 
of  one  hundred,  who  were  inoculated  in  the  presence  of  magistrates 
and  many  medical  men.  This  experiment  was  repeated  with  success. 
Variolous  lymph  was  taken  from  two  lads  at  school,  and  in  ten 


SMALL-POX.  289 

cases  produced  appearances  with  a  perfect  similarity  to  ordinary 
vaccination. 

Thiele  produced  a  benign  vesicle  in  the  following  manner. 
Variolous  lymph  was  diluted  with  warm  cow's  milk,  and  inoculated 
like  ordinary  vaccine  lymph.  Large  vesicles  resulted.  There  were 
febrile  symptoms  from  the  third  to  the  fourth  day,  and  a  secondary 
onset  of  fever  much  more  pronounced  between  the  eleventh  and 
fourteenth  days.  The  areola  was  strongly  marked,  and  not  con- 
fined to  the  inoculated  place,  which  was  occasionally  surrounded 
by  minute  secondary  vesicles.  After  watching  through  ten  removes, 
the  vesicles  finally  assumed  the  characters  of  an  ordinary  vaccination 
with  cow-pox  lymph.  As  soon  as  the  secondary  fever  ceased  to 
occur  inoculation  was  practised  from  arm  to  arm  without  diluting 
the  lymph  with  cow's  milk.  The  lymph  was  designated  lacto-varioline, 
and  the  result  was  variolation  in  its  mildest  form.  The  result  of 
variolating  the  cow  will  be  discussed  in  another  chapter. 

Small-pox  inoculation,  or  variolation,  protected  the  individual 
when  genuine  small-pox  was  produced,  and  .endangered  the  com- 
munity. Persons  inoculated  became  centres  of  infection,  and  con- 
veyed the  disease  to  others.  Haygarth,  although  in  favour  of 
inoculation,  strongly  condemned  its  use  without  precautions  to 
prevent  the  spread  of  the  disease.  "  The  most  serious  and  solid 
objection,"  he  wrote,  "  that  has  been  advanced  against  inoculation 
is  deduced  from  a  comparison  of  the  Bills  of  Mortality  for  a  series 
of  years  in  various  places.  They  show  that  a  larger  proportion 
of  inhabitants  have  died  of  the  small-pox  in  towns  where  it  is  prac- 
tised than  in  the  same  before  it  was  known,  or  in  others  where  it  is 
prohibited." 

Even  Dr.  Dimsdale,  an  ardent  inoculator,  admitted  that  more 
lives  were  lost  in  London  than  before  inoculation  commenced,  and  the 
practice  was  more  detrimental  than  beneficial  to  society ;  and  he 
added  :  "  The  disease  by  general  inoculation  throughout  London 
spreads  by  visitors,  strangers,  servants,  washerwomen,  doctors,  and 
inoculators,  by  means  of  hackney  coaches  in  which  the  sick  are  sent 
out  to  take  the  air,  or  by  sound  persons  approaching  them  in  the 
streets.  The  poor  in  London  are  miserably  lodged  ;  their  habitations 
are  in  close  alleys,  courts,  lanes,  and  old  dirty  houses  ;  they  are  often 
in  want  of  necessaries,  even  of  bedding.  The  fathers  and  mothers 
are  employed  constantly  in  laborious  occupation  abroad,  and  cannot 
attend  the  inoculated  sick."  In  1798  Jenner,  who  had  practised 
small-pox  inoculation,  proposed  the  use  of  a  benign  non-infectious 
lymph  obtained  from  a  disease  of  the  cow  or  horse  as  a  substitute 

19 


290  INFECTIVE    DISEASES. 

for  variolous  lymph,  and  in  1840  small-pox  inoculation  was  prohibited 
by  Act  of  Parliament. 

Stamping-out  System. — The  disappointing  and  dangerous 
results  of  small-pox  inoculation  led  to  a  widespread  demand  for 
some  new  method  for  dealing  with  small-pox.  This  induced  Haygarth 
to  turn  his  attention  to  the  subject,  arid  towards  the  end  of  the 
eighteenth  century  to  bring  before  the  medical  profession  and  the 
public  a  plan  for  stamping  out  the  disease.  Haygarth,  who  was  a 
close  observer  and  an  able  physician,  studied  the  question  of  the 
communicability  of  the  disease  from  one  person  to  another,  and  its 
conveyance  by  infected  clothing  and  other  means,  and  ultimately 
drew  up  rules  and  regulations  for  its  prevention,  the  importance  of 
which  we  are  only  now  beginning  to  fully  acknowledge.  Haygarth's 
essential  doctrine  was  "  that  mankind  was  not  necessarily  subject  to 
the  small-pox,  and  that  it  was  always  caught  by  infection  from  a 
patient  or  the  poisonous  matter,"  and  might  be  avoided  by  observing 
his  Rules  of  Prevention. 

These  rules  comprised  a  regular  system  of  notification  and  isolation. 
Inspectors  were  to  be  provided  to  report  cases  of  small-pox,  and  people 
were  to  be  rewarded  for  carrying  out  the  instructions.  Several  examples 
were  given  of  the  results  at  Chester,  where  the  plan  was  adopted. 

Haygarth  met  with  considerable  encouragement  from  some  of  the 
leaders  of  the  profession.  Dr.  Fothergill  wrote  to  him  in  1778,  saying, 
"  I  have  mentioned  the  intention  of  freeing  this  country  from  the 
small-pox  to  divers  of  the  faculty,  and  shall  continue  to  do  so  as  it  falls 
in  my  way.  The  proposal  is  variously  received,  but  in  exact  proportion 
to  their  humanity." 

In  1793  Haygarth  made  considerable  addition  to  his  rules,  and  urged 
that  legislation  should  follow  to  make  them  compulsory.  Provision  was 
to  be  made  to  reward  the  poor  for  observing  the  rules,  and  public  thanks 
to  the  wealthy  for  their  support  were  to  be  published  in  the  parish  church 
and  newspapers.  Transgression  of  the  rules  was  to  be  punished  by  a  fine  of 
from  £10  to  £50,  one  half  to  go  to  the  informer  and  the  other  half  to  the 
fund  which  supplied  the  expense  of  rewards  to  the  poor,  and  all  details  were 
to  be  supplied  to  the  press.  It  was  further  suggested  that  Great  Britain 
should  be  divided  iuto  districts,  including  a  certain  number  of  parishes 
or  townships,  and  that  to  each  of  them  a  surgeon  or  apothecary  should 
be  appointed  as  inspector  to  see  that  the  regulations  were  exactly 
observed.  In  addition,  there  were  to  be  directors  of  inspectors,  superin- 
tended by  a  commission  of  Physicians  in  London  and  in  Edinburgh.  All 
salaries  were  to  be  paid  by  the  county  rates,  and  the  rewards  for  observing 
the  rules  of  prevention  were  to  be  guaranteed  out  of  the  parish  funds. 
On  the  requisition  of  the  director  and  inspector  of  a  circuit,  power 
was  to  be  given  to  two  or  more  justices  of  the  peace  to  appoint  a  separate 
house  for  the  reception  of  patients  with  the  small-pox.  In  conclusion, 


SMALL- POX.  291 

Haygarth  maintained  that  the  plague  had  been  completely  exterminated 
from  this  country,  for  above  a  century,  by  civil  regulations,  and  that 
there  seemed  to  be  little  doubt  that  the  small-pox  was  propagated  on 
principles  similar  to  the  plague,  and  that  it  also  might  certainly  be 
exterminated  from  this  island. 

Haygarth's  teachings  had  a  profound  influence  upon  both  the  profession 
and  the  educated  public,  but  his  system  of  compulsory  notification  was 
never  carried  out,  for  no  legislation  followed  to  enforce  his  recommenda- 
tions. This  is  a  matter  deeply  to  be  regretted,  for  towards  the  end  of  the 
eighteenth  century  small-pox  was  declining  in  London,  general  sanitation 
was  making  rapid  advances,  small-pox  inoculation,  which  created  fresh 
centres  of  infection,  was  falling  into  disfavour,  small-pox  hospitals  were 
built,  which  served  to  limit  centres  of  infection,  and  the  profession  and 
the  public  were  influenced  by  the  teaching  of  Haygarth  with  regard  to 
the  various  ways  of  avoiding  the  spread  of  the  disease. 

It  only  required  the  compulsory  adoption  of  Haygarth's  system 
uniformly  all  over  the  country  to  have  kept  the  disease  in  control, 
if  not  to  have  entirely  extirpated  it  from  Great  Britain.  That  a 
similar  conviction  existed  at  the  time  is  evidenced  by  an  article  which 
appeared  in  1779  in  the  Medical  and  Chirurgical  Review,  in  which 
the  following  statements  were  made  : — 

k<  Plans  for  the  extirpation  of  the  small-pox  have  been  suggested.  .  .  . 
To  do  this,  however,  the  exertions  of  the  physician  are  incompetent  unless 
they  be  aided  by  the  powerful  hand  of  Governments,  but  this  has  hitherto 
been  withheld.  The  grand  means,  however,  of  extirpating  this  destructive 
malady  is  an  early  and  strict  separation  of  the  infected  from  those  that 
are  sound." 

Small-pox  in  the  present  century  has  been  largely  controlled  by 
legislation,  especially  in  recent  years,  by  the  Public  Health  Acts  for 
England  and  Wales,  for  Scotland,  and  for  Ireland ;  the  Epidemic  and 
other  Diseases  Prevention  Act ;  the  Public  Health  Amendment  Act ; 
the  Labouring  Classes'  Dwellings  Acts  ;  the  Housing  of  the  Working 
Classes  Act ;  the  Public  Health  (Ships)  Act ;  the  Local  Government 
Board  Act — and  various  orders  and  memoranda  of  the  Local  Govern- 
ment Board  ;  the  Infectious  Diseases  Notification  Act ;  the  Infectious 
Diseases  Prevention  Act ;  and  the  Public  Health  (London)  Act. 

By  the  Public  Health  Act  of  1875  England  was  divided  into 
Urban  and  Rural  Sanitary  Districts,  and  powers  were  given  to 
enforce  regulations  of  the  Local  Government  Board  for  guarding 
against  the  spreading  of  infectious  diseases ;  to  provide  medical  aid 
and  accommodation  for  infected  persons,  to  promote  cleansing, 
ventilation,  and  disinfection,  to  provide  hospitals,  to  provide  for 


292  INFECTIVE   DISEASES. 

destruction  or  disinfection  of  infected  bedding,  clothing,  and  other 
articles,  and  to  appoint  Medical  Officers  of  Health. 

As  to  the  value  of  notification  and  isolation  in  cities  such  as 
London  we  have  the  evidence  of  the  Metropolitan  Asylums  Board. 
In  their  Report  for  1889  we  read  in  reference  to  the  diminution  of 
small-pox :  "  These  very  satisfactory  results  confirm  the  view  taken 
by  the  Committee  two  years  ago  to  the  effect  that  the  rapid  and 
systematic  removal  from  crowded  districts  of  infected  persons,  each 
of  whom  might  have  become  a  centre  of  contagion,  is  an  important 
factor  in  stamping  out  small-pox  from  the  metropolitan  population. 
The  notification  of  cases  will  also  greatly  facilitate  the  action  of  the 
managers  in  this  direction." 

More  recently  there  has  been  a  most  striking  confirmation  of 
these  statements.  An  outbreak  of  small-pox  occurred  in  Maryle- 
bone,  and  by  the  energy  of  the  officials  of  the  Board  this  outbreak 
was  suppressed  in  a  few  days  by  means  of  notification  and 
immediate  isolation. 

The  Isolation  Hospitals  Act  of  1893  gives  power  to  County 
Councils  to  provide,  or  cause  to  be  provided,  an  isolation  hospital 
in  any  district  within  their  county.  An  application  to  a  County 
Council  for  the  establishment  of  an  isolation  hospital  may  be  made 
by  any  one  or  more  of  the  authorities  defined  as  local  authorities 
having  jurisdiction  in  the  county  or  any  part  of  the  county. 
Further,  the  County  Council  may  direct  an  inquiry  to  be  made  by 
two  medical  officers  of  health  in  the  county  as  to  the  necessity  of  an 
isolation  hospital  being  established  for  the  use  of  the  inhabitants 
of  any  particular  district  in  the  county,  and  in  the  event  of  such 
medical  officers  reporting  that  such  a  hospital  ought  to  be 
established  for  the  use  of  the  inhabitants  of  a  district,  may  take  the 
same  proceedings  in  all  respects  for  the  establishment  of  such 
hospital,  as  if  a  petition  had  been  presented  by  a  local  authority  for 
the  establishment  of  an  isolation  hospital  for  the  district  named  in 
the  report  of  such  medical  officers  of  health. 

Lastly,  the  Local  Government  Act  of  1894  provides  for  the 
formation  of  District  Councils ;  and  the  powers,  duties,  and 
liabilities  are  principally  those  which  were  conferred  by  the  Public 
Health  Act  of  1875. 

In  the  opinion  of  the  author  the  Government  of  this  country  should 
enter  into  friendly  negotiations  with  the  Governments  of  other  countries, 
so  that  there  might  be  concerted  action  to  prevent  an  avoidable 
disease  like  small-pox.  Much  good  might  result  from  the  formation  of 
a  permanent  International  Board  of  Health.  If  civilisation  is  not  yet 


CATTLE   PLAGUE.  293 

sufficiently  advanced  to  admit  of  a  system  of  international  notification, 
our  Consular  authorities  should  be  instructed  to  give  immediate  notifica 
tion  of  the  existence  of  small-pox  in  other  countries,  and  every  measure 
should  be  enforced  to  diminish  the  possibilities  of  importation.  The 
duties  of  a  Central  Health  Office,  presided  over  by  a  Minister  of  Health, 
should  include  the  collection  of  information  as  to  the  existence  of  small- 
pox in  other  countries,  and  details  should  be  published  in  the  Annual 
Reports  of  the  Department.  Regulations,  for  example,  for  dealing  with 
the  importation  of  rags  from  small-pox  stricken  places  should  be  enforced, 
as  in  the  case  of  cholera  ;  and  if,  in  spite  of  these  precautions,  isolated 
cases  occurred  in  this  country,  they  should  be  dealt  with  promptly. 

Notification  shopld  be  enforced  uniformly  all  over  the  country,  and 
there  is  not  the  slightest  reason  why  the  authorities  and  the  public  should 
not  immediately  receive  information  of  the  existence  of  small-pox,  whilst 
to  procure  immediate  isolation  we  have  only  to  imitate  the  excellent 
ambulance  system  of  the  Metropolitan  Asylums  Board.  To  procure 
prompt  notification  there  must  be  no  loophole  for  evading  the  Act,  and 
there  should  be  a  heavy  penalty  for  failure  to  notify  not  only  small-pox, 
but  any  caxe  which  may  reasonably  be  supposed  to  be  one  of  small-pox. 

The  police  should  be  required  to  report  any  case  of  small-pox  in 
common  lodging-houses  or  shelters  ;  they  should  have  power  to  require 
any  tramp  suffering  from  small-pox,  or  from  any  disease  which  may 
reasonably  be  supposed  to  be  small-pox,  to  be  examined  by  the  medical 
officer  of  the  Union,  and  kept  under  observation,  or  transferred  at  once 
to  the  isolation  hospital  ;  and  inmates  of  the  workhouse  should  be  daily 
inspected,  and  no  case -allowed  to  leave  when  there  is  the  least  suspicion 
of  small-pox  infection. 

Objections  no  doubt  will  be  raised  to  this  proposal,  but  the  frequency 
with  which  small-pox  is  spread  by  tramps  fully  justifies  these  measures. 
All  these  measures  should  be  carried  out  as  a  matter  of  routine,  and 
without  the  semblance  of  panic. 

Isolation  should  be  uniformly  enforced  all  over  the  country,  and 
vaccination  should  be  relegated  to  the  position  of  a  voluntary  auxiliary 
measure,  which  should  never  be  allowed  to  take  the  place  of  sanitary 
regulations  to  stamp  out  the  disease. 

CATTLE  PLAGUE. 

Cattle  plague  is  a  highly  contagious  disease  of  bo  vines  producing 
high  fever,  and  characterised  by  an  eruption  with  a  resemblance  to 
human  small-pox.  The  disease  is  transmissible  to  other  ruminants, 
and  is  inoculable  in  man.  One  attack  gives  immunity  against 
future  attacks.  Cattle  plague  and  small-pox  are  not  intercom- 
municate, and  are  specifically  distinct  diseases,  but  the  resemblance 
between  them  was  recognised  from  early  times.  Ramazzini  published 
an  account  of  the  cattle  pest  in  Italy  in  1711,  and  described  the 
pustules  which  broke  out  over  the  body  as  similar  to  those  of  variola  in 


294  INFECTIVE   DISEASES. 

kind  and  appearance.  Dr.  Layard,  in  1780,  described  this  disease  of 
horned  cattle  as  an  eruptive  fever  of  the  variolous  kind,  with  the 
appearance  and  stages  of  small-pox.  This  resemblance  was  endorsed 
by  Murchison,  one  of  the  Commissioners  appointed  in  1866  to 
inquire  into  the  origin  and  nature  of  cattle  plague. 

Murchison  pointed  out  that  in  both  diseases  the  eruption  con- 
sisted of  pustules  and  scabs,  and  that  in  both  it  extended  from 
the  skin  to  the  interior  of  the  mouth  and  nostrils ;  in  both,  the 
pustules  and  scabs  were  preceded  or  accompanied  by  patches  of 
roseola  ;  in  both,  they  were  occasionally  interspersed  with  petechise  ; 
and  in  both,  they  sometimes  left  behind  pitted  scars  and  discolora- 
tions  on  the  cutis.  The  other  prominent  symptoms  of  rinderpest 
were  also  those  of  small-pox — viz.,  pyrexia,  lumbar  pain,  salivation, 
and  running  from  the  nostrils,  alvine  flux,  albuminuria,  hsematuria, 
and  "  the  typhoid  state."  The  anatomical  lesions  of  the  internal 
organs  in  rinderpest  and  unmodified  small-pox  were  identical — viz., 
congestion  or  inflammation  of  the  mucous  membranes  of  the  air 
passages  and  digestive  canal,  patches  of  ecchymosis  and  even 
gangrene  of  the  stomach  and  other  mucous  surfaces,  and  darkly 
coloured  blood.  In  both  rinderpest  and  small-pox  the  duration 
of  the  pyrexial  stage  was  on  an  average  about  eight  days.  In 
both  diseases  a  peculiarly  offensive  odour  was  exhaled  from  the 
body  before  and  after  death.  The  two  diseases  resembled  one 
another  in  their  extreme  contagiousness,  and  in  the  facility  with 
which  the  poison  was  transmitted  by  foinites.  Both  diseases  were 
easily  propagated  by  inoculation,  and  in  both  cases  the  inoculated 
disease  was  milder  and  less  fatal  than  that  resulting  from  infection. 
In  both  diseases  there  was  a  period  of  incubation,  which  is  shorter 
when  the  poison  has  been  introduced  by  inoculation  than  when  it 
has  been  received  by  infection. 

Ceely  described  the  result  of  an  accidental  inoculation  of  cattle- 
plague  virus  in  the  human  subject.  A  vesicle  was  produced  which 
so  closely  corresponded  with  the  result  of  inoculated  cow-pox  that 
Ceely  inclined  to  the  belief  that  cattle  plague  was  a  malignant  form 
of  cow-pox.  The  following  is  the  account  of  this  case  as  reported 
by  Ceely.  Mr.  Hancock,  a  veterinary  inspector  at  Uxbridge,  was 
engaged  in  superintending  the  autopsy  of  a  bullock  recently  dead  of 
cattle  plague.  His  assistant,  who  was  performing  the  operation, 
while  occupied  in  removing  the  skin  from  the  scrotum,  accidentally 
punctured  the  back  of  Mr.  Hancock's  hand  with  the  point  of  the 
knife.  The  puncture  being  slight  was  disregarded  at  the  time,  but 
was  washed  as  soon  as  practicable,  and  thought  of  no  more.  Five 


CATTLE   PLAGUE.  295 

days  afterwards,  a  small,  slightly  elevated,  hard  pimple  was  felt 
and  seen  on  the  site  of  the  puncture.  This  gradually  advanced 
till  the  ninth  day  of  the  puncture,  the  fourth  from  papulatipn, 
when  the  enlargement  became  distinctly  vesicular.  At  that  time 
there  were  but  slight  constitutional  symptoms.  On  the  next  day, 
the  tenth  from  the  receipt  of  the  puncture,  the  fifth  from  papula tion, 
and  the  second  from  vesiculation,  Mr.  Hancock  consulted  Mr.  Rayner, 
of  Uxbridge,  who,  on  seeing  the  hand,  inquired  if  the  patient  had 
been  handling  the  udder  of  a  cow,  as  he  thought  he  could  recognise 
a  cow-pox  vesicle  of  the  ninth  day.  The  vesicle  was  distended 
with  thin  lymph,  its  margin  elevated  and  slightly  brown,  its  centre 
depressed  and  brownish,  and  the  whole  surrounded  with  a  large 
bright  red  areola.  There  was  then  considerable  tumefaction  extend- 
ing from  the  knuckles  to  above  the  wrist.  The  absorbent  vessels 
were  considerably  inflamed,  and,  like  the  axillary  glands,  were  tender 
and  painful ;  the  pulse,  naturally  slow,  was  accelerated ;  there  was 
much  pain  in  the  back  and  limbs,  severe  distracting  headache,  etc. ; 
all  of  which  symptoms  continued  to  increase  during  the  two  following 
days.  At  the  end  of  that  time  the  diffused  areola  had  extended  as 
far  as  the  elbow.  Fifteen  days  after  the  puncture,  and  ten  days  after 
papulation,  the  local  inflammation  and  constitutional  symptoms  had 
partially  subsided.  The  vesicle  contained  a  rather  turbid  brownish 
fluid,  and  there  were  present  all  the  indications  of  a  declining  vaccine 
vesicle. 

Murchison  also  saw  and  described  the  case,  and  gave  practically 
the  same  account  of  it.  He  pointed  out  that  the  appearances  and 
the  entire  history  were  very  different  from  the  results  of  a  poisoned 
wound,  but  coincided  with  the  appearances  seen  after  vaccination. 

In  1832  Macpherson,  in  Bengal,  inoculated  eleven  native  children 
with  cattle- plague  crusts.  There  was  no  result  in  six,  others 
suffered  from  local  inflammation,  and  in  one  a  vesicle  formed. 
With  lymph  from  this  vesicle  other  children  were  inoculated.  The 
results  in  all  were  similar  in  appearance  to  those  of  vaccination. 
Two  children  were  subsequently  inoculated  with  human  variola,  and 
were  said  to  be  protected. 

In  1834  Macpherson's  example  was  followed  by  Mr.  Furiiell 
in  Assam.  Furnell  inoculated  four  children  with  cattle-plague 
crusts  without  result,  but  his  assistant  succeeded  with  crusts  taken 
from  the  back  and  abdomen  of  the  diseased  cattle,  and  carried  on 
the  lymph  from  child  to  child.  In  one  case  there  was  a  general 
eruption.  Furnell  inoculated  his  own  child  from  one  of  the  native 
children :  a  copious  eruption  followed,  and  the  child  died.  Furnell 


296  INFECTIVE    DISEASES. 

after  this  misfortune  issued  a  strong  warning  against  taking  the 
virus  from  the  cow.  The  experiments  were  made  in  the  belief  that 
cattle  plague  was  really  small-pox  in  cattle,  and  that  the  virus 
would  protect  against  human  variola. 

Similar    results    were    obtained    by    Mr.    Wood    at    Gowalpara 
in  1838. 

Bacteria  in  Cattle  Plague. — Semner  cultivated  streptococci 
from  the  blood  and  lymphatic  glands  of  a  sheep  suffering  from 
cattle  plague.  A  calf  inoculated  with  a  cultivation  died  in  seven 
days.  The  cocci  were  stated  to  lose  their  virulence  by  cultivation, 
and  the  weakened  cultivation  to  protect  against  the  virulent  disease. 
The  micro-organism  was  very  probably  Streptococcus  pyogenes, 
and  the  calf  may  have  died  of  septic  infection.  There  can  be  110 
doubt  that  the  nature  of  the  contagium  of  cattle  plague  is  unknown. 
Protective  Inoculation. — In  the  great  epidemic  of  cattle 
plague  in  England  in  1866,  owing  to  a  belief  that  the  analogy 
between  cattle  plague  and  small-pox  was  closer  than  it  really  is, 
vaccination  with  cow-pox  was  attempted  as  a  preventive  measure, 
but  was  proved  to  be  absolutely  useless. 

Stamping-out  System. — When  cattle  plague  was  imported  in 
1865  into  London,  dairymen  and  stock-owners  made  no  attempts 
to  prevent  the  extension  of  the  disease,  so  that  it  spread  rapidly 
all  over  the  country  through  disposal  of  infected  cattle.  The  losses 
were  enormous,  and  an  Order  in  Council  was  passed  in  July  1865, 
directing  dairymen  and  others  to  notify  outbreaks  of  any  contagious 
or  infectious  disease  among  the  animals  under  their  charge.  A 
Veterinary  Department  of  the  State  was  formed,  and  inspectors 
appointed  in  various  parts  of  the  country.  A  short  Act  was  passed 
in  February  1866.  A  stamping-out  system,  consisting  of  compulsory 
notification  and  the  slaughter  of  diseased  animals,  was  soon  brought 
to  the  notice  of  the  public.  There  was  violent  opposition,  but 
nevertheless,  after  some  delay,  the  system  was  carried  out.  The 
number  of  cases  of  cattle  plague  had  reached  18,000  weekly,  and 
on  the  introduction  of  the  stamping-out  system  the  disease  rapidly 
declined.  The  disease  was  again  imported  into  Great  Britain  in 
1872,  and  there  were  outbreaks  in  1877.  In  each  instance  the 
disease  was  promptly  stamped  out,  and  ever  since  that  year  the 
disease  has  been  kept  out  of  this  country. 


CHAPTER   XXI. 

SHEEP-POX. — FOOT-AND-MOUTH   DISEASE. 

SHEEP-POX. 

SHEEP-POX,  or  variola  ovina,  is  an  acute  febrile  disease  accompanied 
by  a  general  vesiculo-pustular  eruption,  highly  infectious,  and 
capable  of  being  propagated  by  inoculation  or  clavelisation.  It  is 
a  common  disease  in  some  parts  of  Europe.  In  France  the  disease 
is  called  la  clavelee,  and  in  Italy  vaccuolo.  It  has  been  introduced 
on  several  occasions  into  this  country,  but  has  been  effectually 
stamped  out.  As  in  human  small-pox,  there  are  varieties — the 
benign  and  the  malignant,  the  discrete  and  the  confluent ;  and  one 
attack  is  protective  against  the  disease  in  future. 

It  is  very  closely  analogous  to  human  small-pox.  Vaccination 
with  cow-pox  lymph  has  been  employed  to  protect  sheep  from  sheep- 
pox,  but  unsuccessfully,  and  lymph  for  vaccination  has  been  raised 
from  sheep-pox  to  protect  human  beings  from  small-pox.  These 
experiments  were  first  performed  in  Italy. 

Marchelli,  in  1802,  took  lymph  from  the  vesicles  of  sheep-pox,  and 
inoculated  children.  Sacco  repeated  these  experiments,  and  found 
there  was  no  appreciable  difference  from  the  results  obtained  with 
cow-pox  lymph.  Dr.  Legni  carried  on  the  inoculations  with  ovine 
virus  from  arm  to  arm  for  several  years,  and  when  small-pox 
occurred  in  Pesaro,  it  was  said  that  all  those  who  were  inoculated 
with  the  sheep  virus  were  protected. 

Inoculation  of  children  with  ovine  virus,  direct  from  sheep,  was 
repeated  by  Sacco  and  Magnani  in  1806. 

Marson  in  England  succeeded  in  producing  on  the  human 
subject  a  vesicle  with  the  physical  characters  of  the  vaccine  vesicle. 
The  vesicle  had  a  bluer  tinge,  and  subsequent  inoculation  of  the 
patient  with  human  variola  was  ineffectual.  Other  experimenters 
were  unsuccessful,  but  their  failures,  as  in  the  case  of  variolation  of 
the  cow,  do  not  invalidate  the  results  of  those  who  were  successful. 

297 


298  INFECTIVE   DISEASES. 

Sheep-pox  and  cow-pox  are  quite  distinct  diseases.  Sheep-pox 
is  highly  infectious,  whereas  cow-pox  is  only  conveyed  by  direct 
inoculation,  and  is  never  infectious,  and  further,  cow-pox  inoculated 
in  sheep  does  not  produce  sheep- pox. 

Bacteria  in  Sheep-pox. — Hallier  and  Zurn,  Klein,  and  others, 
have  found  micrococci  and  bacteria  in  the  lymph  of  the  vesicles  of 
sheep-pox,  but  they  are  only  accidental  epiphytes.  The  nature  of 
the  contagium  is  unknown. 

Protective  Inoculation. — Extensive  experiments  were  carried 
out  in  England  to  test  the  protective  power  of  vaccination  against 
sheep-pox.  According  to  Marson  and  Simmonds,  it  was  very  difficult 
to  get  cow-pox  to  take  on  sheep,  and  when  an  effect  was  produced, 
the  resulting  affection,  even  when  developed  to  its  fullest  extent,  was 
very  unlike  the  same  disease  in  the  human  subject.  In  the  sheep 
it  seldom  produced  anything  more  than  a  small  papule,  which  occa- 
sionally resulted  in  the  formation  of  a  minute  vesicle,  or  more 
commonly,  a  pustule,  which  was  sometimes,  although  very  rarely, 
surrounded  by  a  slight  areola.  Generally,  however,  neither  vesica- 
tion  nor  pustulation  followed,  but  a  small  scab  was  produced,  which 
soon  fell  from  the  site  of  the  puncture,  leaving  no  trace  behind.  The 
disease  passed  quickly  and  irregularly  through  its  several  stages, 
and  terminated  by  the  eighth  or  ninth  day,  and  not  unfrequently 
even  before  that  time.  Lymph  was  but  rarely  obtainable,  and  then 
only  in  the  smallest  quantity,  and  this  on  the  fifth  or  sixth  day  suc- 
ceeding the  vaccination.  The  effects  were  only  local,  and  the  animal's 
health  was  not  impaired. 

Sheep  were  found  to  be  just  as  susceptible  of  the  cow-pox  virus 
on  subsequent  repetition  of  the  inoculation  as  they  were  in  the 
first  instance,  and  hence  the  conclusion  that  cow-pox  was  utterly 
worthless  as  a  protective  against  sheep-pox.  According  to  Depaulr 
however,  cow-pox  takes  characteristically  on  sheep,  and  sheep- pox 
lymph  inoculated  on  cows  produces  a  result  indistinguishable  from 
the  appearances  obtained  with  the  inoculation  of  cow-pox  lymph, 
It  is  impossible  to  say  whether  these  conflicting  results  depended 
upon  the  employment  in  the  experiments  of  different  breeds  of  sheep 
or  different  stocks  of  vaccine  lymph. 

The  objection  to  clavelisation  or  ovination  is  that  the  disease 
may  be  introduced  in  localities  where  it  was  previously  unknown. 
By  ovination,  as  in  the  analogous  case  of  variolation,  fresh  centres 
of  infection  are  created,  whereas  every  precaution  should  be  taken 
to  prevent  the  introduction  of  the  disease. 

Stamping-out  System. — Sheep-pox  has  been  imported  into  this 


FOOT-AND-MOUTH   DISEASE.  299 

country  on  several  occasions.  It  was  introduced  in  1847,  and  again 
in  1862;  in  1865  it  was  introduced  again,  and  active  measures  of 
repression  were  at  once  taken.  The  diseased  flocks  were  carefully 
isolated,  and  day  by  day  as  fresh  cases  occurred  the  diseased  animals 
were  killed  and  buried.  Owing  to  the  adoption  of  these  precaution- 
ary measures,  the  affection  did  not  extend  beyond  the  flock  among 
which  it  first  appeared.  It  was  introduced  again  in  1866  at  Long 
Buckby,  in  Northamptonshire.  In  this  case  the  disease  was  exter- 
minated by  the  slaughter  and  burial  of  the  whole  flock,  and  imme- 
diate application  of  disinfectants  to  the  hurdles  and  other  things  with 
which  the  sheep  had  been  in  contact.  Then  it  was  introduced 
again  in  Cheshire,  and  strict  isolation  being  enforced  the  infection 
died  out.  Since  1866  we  have  had  no  outbreak  of  sheep-pox  in 
this  kingdom,  but  foreign  sheep  have  been  landed  with  sheep-pox 
in  1868,  1869,  1870,  1871,  1875,  1876,  1878,  and  1880,  but  the 
disease  has  been  prevented  from  spreading. 

The  Sheep-pox  Order  of  1895  provides  for  the  notification  of 
the  disease,  for  disinfection  and  for  compulsory  slaughter  of  infected 
sheep,  and  prohibits  the  movement  of  diseased  or  suspected  sheep, 
and  the  local  authority  may,  if  they  think  fit,  order  the  slaughter 
of  suspected  sheep  and  of  sheep  which  have  been  in  contact  with 
diseased  sheep. 

FOOT-AND-MOUTH  DISEASE. 

Foot-and-mouth  disease  is  a  highly  contagious  and  infectious 
febrile  disease,  characterised  by  a  vesicular  eruption  affecting  the 
lips,  tongue,  roof  of  the  mouth,  and  feet  of  sheep,  cattle,  and  pigs, 
and  according  to  some  observers  it  also  attacks  horses,  poultry, 
hares,  and  rabbits.  Sometimes  the  mouth  only  is  affected,  in  other 
cases  the  principal  seat  of  the  eruption  is  in  the  feet.  The  vesicles 
soon  break  and  give  rise  to  ulcers.  When  these  occur  in  the  mouth 
they  cause  pain  and  difficulty  in  taking  food.  Extensive  ulceration 
may  occur  on  the  feet,  causing  great  pain  and  lameness.  In  milch 
cows  it  sometimes  happens  that  the  eruption  occurs  on  the  udder  and 
teats,  and  it  is  this  manifestation  of  the  disease  which  has  received 
so  much  attention  from  Eayer.  The  milk  is  contaminated  by  the 
discharge  of  the  vesicles,  and  is  unfit  for  use,  either  as  food  for  the 
human  being  or  for  the  lower  animals.  It  induces  a  vesicular 
eruption  in  the  mouth,  larynx,  pharynx,  and  intestinal  canal.  It 
acts  most  vigorously  when  administered  warm  to  young  animals, 
and  calves  occasionally  die  quite  suddenly  after  sucking  cows 


300  INFECTIVE    DISEASES. 

affected  with  the  eruption  on  the  teats.     Fatal  effects  also  result 
when  the  milk  is  administered  to  young  pigs. 

It  has  been  stated  that  no  injurious  consequences  arise  from  the 
consumption  of  the  milk  by  human  beings,  but  there  is  abundant 
evidence  to  the  contrary,  and  the  conflicting  opinions  probably  arise 
from  the  fact  that  milk  is  seldom  drunk  direct  from  the  cow,  and 
rarely  in  an  undiluted  form.  Hertwig  experimented  upon  himself 
with  milk  freshly  drawn  from  a  cow  with  the  eruption.  He  drank 
a  pint,  and  two  days  afterwards  experienced  slight  fever,  restless- 
ness, and  headache.  The  mouth  was  dry  and  hot,  and  there  was 
tingling  in  the  skin  of  the  hands  and  fingers.  These  symptoms 
continued  for  seven  days  after  taking  the  milk.  On  the  ninth  day 
vesicles  had  formed  on  the  tongue,  principally  on  the  edges,  and  011 
the  mucous  membrane  of  the  cheeks  and  lips  (the  largest  being 
about  the  size  of  a  lentil).  They  were  yellowish- white  in  colour,  and 
contained  a  whitish  turbid  liquid,  which  flowed  when  the  vesicles 
were  pricked  with  a  needle.  At  the  same  time  a  number  of  vesicles- 
developed  on  the  hands  arid  fingers  ;  and  most  of  them  at  the 
time  of  their  first  appearance  were  the  size  of  a  millet  seed.  They 
were  firm  to  the  touch,  yellowish -white,  and  occasioned  a  slight 
tingling.  The  vesicles  of  the  mouth  increased  in  size  and  eventually 
broke,  and  the  epithelium  detached  itself  completely  from  the  affected 
parts,  leaving  dark  red  spots,  which  disappeared  gradually.  The 
slight  fever  present  during  the  first  days  ceased  after  the  appearance 
of  the  eruption ;  but  from  this  time,  until  the  disappearance  of 
the  red  spots,  Hertwig  felt  a  continual  burning  pain  in  the  mouth, 
and  speaking  and  deglutition  caused  considerable  uneasiness.  On 
the  lips  the  vesicles  dried  up,  and  were  covered  with  thin  brownish 
crusts,  which  fell  off  ten  days  after  the  appearance  of  the  first 
vesicles.  The  vesicles  which  developed  on  the  hands  ran  a  slower 
course.  From  the  tenth  to  the  thirteenth  day  they  filled  with  a 
liquid,  like  turbid  lymph.  They  were  large  and  confluent,  and 
finally  broke  and  dried  up. 

Bacteria  in  Foot-and-mouth  Disease.— Klein  in  1885  isolated 
from  the  vesicles  a  streptococcus  which  in  its  microscopical  and  its 
cultural  characters  on  gelatine,  agar  and  blood  serum  resembled 
Streptococcus  pyogeiies.  Minute  differences  in  the  size  of  the 
colonies  and  in  their  rate  of  growth,  and  in  the  character  of  the 
chains,  were  observed  on  making  comparative  cultures  with 
Streptococcus  pyogenes  from  a  human  source,  but  no  comparison 
was  made  with  Streptococcus  pyogenes  from  acute  suppuration  in 
cattle.  Baumgarten  regarded  this  micro-organism  as  Streptococcus 


FOOT-AND-MOUTH   DISEASE.  301 

pyogenes,  and  not  as  the  contagium  of  the  disease.  The  author  has 
pointed  out  the  variation  which  exists  in  the  size  of  the  chains  and 
of  the  colonies,  and  the  difference  which  is  found  in  the  rate  of 
growth  of  cultures  of  Streptococcus  pyogenes,  and  these  variations  are 
especially  marked  in  Streptococcus  pyogenes  bo  vis.  Klein  believes 
that  the  administrations  of  broth  cultures  produced  the  disease  in 
sheep,  but  the  results  were  very  probably  due  to  accidental  infection. 
It  is  well  known  how  very  readily  foot-and-mouth  disease  is  spread. 
The  appearance  of  a  case  in  a  flock  of  sheep  or  a  herd  of  cattle  will 
be  almost  certain  to  be  followed  by  all  or  nearly  all  of  the  other 
animals  being  infected  with  great  rapidity.  The  virus  clings  to  the 
clothes  of  shepherds  and  others  who  have  been  in  contact  with 
infected  sheep,  and  may  be  readily  conveyed  to  healthy  animals  by 
those  who  have  been  visiting  infected  premises. 

Schottelius  described  chains  composed  of  rounded  elements,  some 
of  which  resembled  an  amoeba  or  plasmodium.  The  chains  were  said 
to  be  motile,  and  delicate  growths  were  obtained  in  blood  serum  and 
agar,  and  in  broth  and  on  potato.  Inoculation  in  sheep  and  pigs 
and  numerous  small  animals  gave  negative  results.  These  organisms 
were  described  as  streptocytes,  to  distinguish  them  from  bacteria. 

Piani  and  Fiorentini  investigated  the  contents  of  the  vesicles,  and 
also  described  corpuscular  elements  exhibiting  amoeboid  movements. 
They  regarded  these  bodies  as  protozoa,  and  concluded  that  foot-and- 
mouth  disease  is  due  to  their  presence. 

Until  a  micro-organism  is  cultivated  which  will  produce  sheep- 
pox  in  sheep  on  a  farm  or  on  premises  where  the  disease  does  not 
exist,  and  where  there  can  be  no  possibility  of  accidental  infection, 
we  are  fully  justified  in  concluding  that  the  nature  of  the  contagium 
of  this  disease  is  unknown. 

Stamping-out  System. — Foot-and-mouth  disease  was  imported 
into  this  country  in  1839.  It  has  been  successfully  dealt  with  by  the 
stamping-out  system,  which  in  this  case  is  very  difficult  to  apply 
because  of  the  very  short  period  of  incubation,  and  the  value  of 
the  stamping-out  method  very  greatly  depends  upon  the  length 
of  the  incubation  period.  Foot-and-mouth  disease  very  often, 
from  infection  to  recovery,  does  not  exceed  ten  days;  yet  accord  in.-: 
to  the  reports  of  the  Board  of  Agriculture,  when  foot-and-mouth 
disease  exists  in  a  manageable  state,  perfect  isolation  and  effectual 
disinfection  have  proved  equal  to  the  complete  control  of  the 
spreading  of  the  infection,  and  the  final  extinction  of  the  disease. 
Nothing  more  is  necessary  in  any  case  than  to  close  up  all  the 
channels  through  which  infected  matter  can  be  conveyed;  but  in 


302  INFECTIVE   DISEASES. 

order  that  this  may  be  done  close  supervision  by  conscientious  and 
responsible  officers  is  required ;  without  it  the  case  is  hopeless. 

The  Foot-and-mouth  Disease  Order  of  1895  enforces  notification, 
isolation,  and  disinfection,  and  the  question  of  slaughter  is  left  to 
the  local  authority. 

(1)  A  local  authority  may,  if  they  think  fit,  cause  to  be  slaughtered — 
(«)  Any  cattle,  sheep,  or  swine  affected  with  foot-and-mouth  disease 

or  suspected  of  being  so  affected  ;  and 

(&)  Any  cattle,  sheep,  or  swine  being  or  having  been  in  the  same  field, 
shed,  or  other  place,  or  in  the  same  herd  or  flock  or  otherwise  in 
contact  with  animals  affected  with  foot-and-mouth  disease,  or  being 
or  having  been,  in  the  opinion  of  the  local  authority,  in  any  way 
exposed  to  the  infection  of  foot-and-mouth  disease. 


CHAPTER    XXII. 

HORSE-POX.—  COW-POX. 

CONSTITUTIONAL  GREASE  OR  HORSE- POX. 

HORSE- POX  is  a  vesicular  disease  of  the  horse  communicable  from 
animal  to  animal  by  inoculation,  but  never  infectious.  It  is 
communicable  by  inoculation  to  man,  and  the  attenuated  virus 
produces  phenomena  indistinguishable  from  the  results  of  vaccina- 
tion with  cow-pox  lymph. 

The  existence  of  this  disease  of  the  horse  had  long  been  known 
to  farmers  and  farriers,  but  Jenner  was  the  first  to  draw  attention 
to  it  in  writing.  "  There  is  a  disease  to  which  the  horse  from  his 
state  of  domestication  is  frequently  subject.  The  farriers  have 
termed  it  the  grease ;  it  is  an  inflammation  and  swelling  in  the  heel 
accompanied  at  its  commencement  with  small  cracks  and  fissures, 
from  which  issues  matter  possessing  properties  of  a  very  peculiar 
kind."  Jenner  gave  several  instances  in  which  this  disease  was 
communicated  to  man  and  to  cows. 

Thus,  a  man  named  Merret  attended  to  some  horses  with  sore 
heels  and  also  milked  the  cows.  The  cows  were  infected,  and  the 
man  had  several  sores  upon  his  hands. 

William  Smith,  on  another  farm,  attended  to  horses  with  sore 
heels  and  milked  the  cows  also.  The  cows  were  infected,  and  on  one 
of  Smith's  hands  there  were  several  ulcerated  sores. 

Simon  Nicholls  applied  dressings  to  the  sore  heels  of  one  of  his 
master's  horses  and  at  the  same  time  milked  the  cows,  and  the  cows 
were  infected  in  consequence. 

A  mare,  the  property  of  a,  dairy  farmer,  had  sore  heels,  and 
was  attended  to  by  the  men  of  the  farm,  Thomas  Yirgoe,  William 
Wherret,  and  William  Haynes.  They  contracted  "sores  on  their 
hands,  followed  by  inflamed  lymphatic  glands  in  the  arms  and 
axillae,  shiverings  succeeded  by  heat,  lassitude,  and  general  pains  in 
the  limbs,"  and  the  disease  was  also  communicated  to  the  cows. 

303 


304  INFECTIVE   DISEASES. 

But  Jenner's  experience  of  this  disease  was  not  limited  to  cases 
in  which  the  eruption  occurred  in  the  heel. 

He  mentions  a  case  in  which — 

"  An  extensive  inflammation  of  the  erysipelatous  kind  appeared 
without  any  cause  upon  the  upper  part  of  the  thigh  of  a  sucking- 
colt.  The  inflammation  continued  several  weeks,  and  at  length 
terminated  in  the  formation  of  three  or  four  small  abscesses."  Those 
who  dressed  the  colt  also  milked  the  cows  on  the  farm,  and  communi- 
cated the  disease  to  them. 

Subsequently,  Jenner  gave  a  more  comprehensive  description  of 
this  disease. 

"The  skin  of  the  horse' is  subject  to  an  eruptive  disease  of  a 
vesicular  character,  which  vesicle  contains  a  limpid  fluid,  showing 
itself  more  commonly  in  the  heels.  The  legs  first  become  cedematous, 
and  then  fissures  are  observed.  The  skin  contiguous  to  these  fissures, 
when  accurately  examined,  is  seen  studded  with  small  vesicles  sur- 
rounded by  an  areola.  These  vesicles  contain  the  specific  fluid.  It 
is  the  ill- management  of  the  horse  in  the  stable  that  occasions  the 
malady  to  appear  more  frequently  in  the  heel  than  in  other  parts. 
I  have  detected  it  connected  with  a  sore  on  the  neck  of  the  horse, 
and  on  the  thigh  of  a  colt." 

Mr.  Moore,  of  Chalford  Hill,  described  a  case  in  1797,  and  re- 
garded the  disease  as  virulent  grease.  His  horse  was  attacked  with 
what  was  supposed  to  be  ordinary  "  grease."  A  cow  was  subse- 
quently infected,  and  the  disease  communicated  to  the  servant,  who 
had  "  eruptions  on  his  hands,  face,  and  many  other  parts  of  the 
body,  the  pustules  appearing  large,  and  not  much  unlike  the  small- 
pox, for  which  he  had  been  inoculated  a  year  and  a  half  before,  and 
had  then  a  very  heavy  burden." 

In  1798,  Mr.  Fewster,  of  Thornbury,  met  with  a  case  of  this 
equine  malady,  and  wrote  a  very  full  account  to  Jenner  of  its 
transmission  to  the  human  subject. 

"  William  Morris,  aged  thirty-two,  servant  to  Mr.  Cox  of 
Almonsbury  in  this  county,  applied  to  me  the  2nd  of  April,  1798. 
He  told  me  that  four  days  before  he  found  a  stiffness  and  swelling 
in  both  his  hands,  which  were  so  painful  it  was  with  difficulty  he 
continued  his  work ;  that  he  had  been  seized  with  pain  in  his  head, 
small  of  the  back,  and  limbs,  and  with  frequent  chilly  fits  succeeded 
by  fever.  On  examination  I  found  him  still  affected  with  these 
symptoms,  and  there  was  great  prostration  of  strength.  Many 
parts  of  his  hands  on  the  inside  were  chapped,  and  on  the  middle 
joint  of  the  thumb  of  the  right  hand  there  was  a  small  phagedaenic 


CONSTITUTIONAL   GREASE   OR   HORSE-POX.  305 

ulcer,  about  the  size  of  "a  large  pea,  discharging  an  ichorous  fluid. 
On  the  middle  finger  of  the  same  hand  there  was  another  ulcer  of  a 
similar  kind.  These  sores  were  of  a  circular  form,  and  he  described 
their  first  appearance  as  being  somewhat  like  blisters  arising  from  a 
burn.  He  complained  of  excessive  pain,  which  extended  up  his  arm 
into  the  axilla.  On  the  5th  of  April  I  again  saw  him,  and  found 
him  still  complaining  of  pain  in  both  his  hands,  nor  were  his  febrile 
symptoms  at  all  relieved.  The  ulcers  had  now  spread  to  the  size  of 
a  seven-shilling  gold  coin,  and  another  ulcer,  which  I  had  not  noticed 
before,  appeared  on  the  first  joint  of  the  forefinger  of  the  left  hand,, 
equally  painful  with  that  on  the  right.  I  ordered  him  to  bathe  hi& 
hands  in  warm  bran  and  water,  apply  escharotics  to  the  ulcers,  and 
wrapped  his  hands  up  in  a  soft  cataplasm.  The  next  day  he  wa& 
much  relieved,  and  in  something  more  than  a  fortnight  got  welL 
He  lost  his  nails  from  the  thumb  and  fingers  that  were  ulcerated." 

Mr.  Tanner,  a  veterinary  surgeon,  was  the  first  to  succeed  in 
experimentally  transmitting  horse- pox  to  the  teats  of  a  cow  by 
inoculating  some  of  the  liquid  matter  from  the  heel  of  a  horse.  From 
handling  the  cow's  teats  he  became  infected  himself,  and  had  two- 
pustules  on  his  hand,  which  brought  on  inflammation,  and  made 
him  unwell  for  several  days.  The  matter  from  the  cow  and  from  his 
own  hand  proved  efficacious  in  infecting  both  human  subjects  and 
cattle. 

In  1801  Dr.  Loy  published  his  experiments.  A  butcher  had 
painful  sores  from  dressing  a  horse  suffering  from  *  grease/  and  Dr. 
Loy  succeeded  in  transmitting  the  disease  to  the  udder  of  a  cow. 
Matter  was  taken  from  the  cow  and  inserted  into  the  arm  of  a  child. 
Dr.  Loy  also  inoculated  a  child  direct  from  a  horse  suffering  from 
*  grease/  and  subsequently  five  other  children  from  this  child. 

From  his  experiments  and  observations  Dr.  Loy  was  led  to 
differentiate  constitutional  grease  from  the  merely  local  affection 
commonly  known  as  the  grease,  and  thus  he  explained  the  failure 
on  the  part  of  many  experimenters  to  transmit  this  disease  to- 
the  cow. 

"  This  fact  induces  me  " — he  says — "  to  suspect  that  two  kinds 
of  grease  exist,  differing  from  each  other  in  the  power  of  giving 
disease  to  the  human  or  brute  animal ;  and  there  is  another  circum- 
stance which  renders  this  supposition  probable.  The  horses  that 
communicated  the  infection  to  their  dressers  were  affected  with  a 
general  as  well  as  a  topical  disease.  The  animals  at  the  commence- 
ment of  their  disease  were  evidently  in  a  feverish  state,  from  which 
they  were  relieved  as  soon  as  the  complaint  appeared  at  their  heels r 

20 


306  INFECTIVE    DISEASES. 

and  an  eruption  upon  the  skin.  The  horse,  too,  from  which  the 
infectious  matter  was  procured  for  inoculation,  had  a  considerable 
indisposition,  previous  to  the  disease  at  his  heels,  which  was  attended, 
as  in  the  others,  with  an  eruption  over  the  greatest  part  of  his 
body;  but  those  that  did  not  communicate  the  disease  at  all,  had 
a  local  affection  only.  From  this  perhaps  may  be  explained  the 
want  of  success  attending  the  experiments  of  the  gentlemen  I  have 
mentioned." 

Experiments  with  horse-pox  were  also  made  about  this  time  on 
the  Continent.  Sacco  made  some  observations  upon  this  disease  at 
Milan.  Several  horses  were  suffering  from  what  was  called  giardoni, 
and  Sacco's  servant  was  attacked  on  both  arms,  from  dressing  one 
of  his  horses  troubled  with  this  disease.  Several  children  and  cows 
were  inoculated  from  the  horses,  but  without  success.  In  another 
instance,  a  coachman  went  to  the  hospital  with  the  eruption  on  his 
hands,  and  the  disease  was  successfully  communicated  to  three  out 
of  nine  children. 

In  1803  Dr.  Marcet  described  some  experiments  which  had  been 
made  at  Salonica  by  M.  La  Font.  The  disease  was  known  to  the 
farriers  in  Macedonia  as  javart.  In  one  case,  a  horse  was  attacked 
with  feverish  symptoms  that  ceased  as  soon  as  the  eruption  appeared. 
The  fore  legs  were  much  swelled  and  several  ulcers  formed.  M.  La 
Font  took  some  of  the  discharge  from  an  ulcer  and  inoculated  a  cow 
and  three  children,  and  succeeded  in  transmitting  the  disease  to  two 
of  the  latter. 

Vaccinogenic  grease  was  observed  in  Paris  in  1812,  and  Baron 
cites  the  case  of  a  coachman  who,  after  dressing  a  horse  with  the 
"  grease,"  had  a  crop  of  pustules  on  his  hands,  from  which  the  disease 
was  experimentally  transmitted  by  inoculation  to  two  children.  A 
series  of  inoculations  was  started  from  an  infant  who  was  infected 
from  one  of  the  scabs  taken  from  the  pustules  on  the  hand  of  the 
coachman. 

In  1813  Mr.  Melon,  a  surgeon  at  Lichfield,  met  with  vaccinogenic 
grease  in  the  horse,  and  some  of  the  virus  was  sent  to  Jenner,  who 
carried  on  a  series  of  arm  to  arm  equinations  for  some  months.  And 
again  in  1817,  vaccinogenic  grease  broke  out  in  a  farm  at  Wansell. 
The  farm-servants  and  the  cows  were  infected,  and  Jenner  employed 
this  equine  matter  for  a  series  of  inoculations  for  eight  months. 

In  1817  Baron  described  a  case  of  a  young  man  who  had  not  less 
than  fifty  pustules  on  his  hands  and  wrists  from  dressing  a  horse  with 
this  disease,  and  in  the  following  year  Baron  obtained  some  fresh 
equine  virus  from  the  hands  of  a  boy  who  had  been  infected  directly 


CONSTITUTIONAL   GREASE   OR   HORSE-POX.  307 

from  a  horse.  The  disease  assumed  a  pustular  form,  and  extended 
over  both  arms. 

In  1818  Kahlert  met  with  this  equine  disease  in  Bohemia,  and 
confirmed  the  experiments  made  by  Loy  and  Sacco.  Kahlert  noticed 
that  the  joint  of  the  foot  was  swollen,  and  moisture  exuded  from  it, 
and  that  the  posterior  part  of  the  pastern  was  slightly  red,  swollen 
and  hotter  than  the  neighbouring  parts,  and  a  clear  yellowish  fluid 
with  a  peculiar  odour  escaped.  At  the  slightest  touch  the  animal 
showed  signs  of  pain  ;  the  hair  was  stuck  together.  The  disease  was 
successfully  transmitted  to  cows  and  from  cows  to  children. 

In  1860  the  horses  at  Bieumes,  near  Toulouse,  were  attacked  by 
an  epizootic  malady  ;  in  less  than  three  weeks  there  were  more  than 
one  hundred  cases.  According  to  the  veterinary  surgeon,  M.  Sarrana, 
the  animals  suffered  from  slight  fever,  rapidly  followed  by  local 
symptoms,  the  most  marked  of  which  were  swelling  of  the  hocks,  and 
an  eruption  of  small  pustules  on  the  surface  of  the  swollen  parts, 
which  -were,  at  the  same  time,  hot  and  painful.  After  three  to  five 
clays  there  was  a  discharge  from  the  pastern  which  continued  for  eight 
to  ten  days,  during  which  the  inflammation  gradually  diminished. 
The  pustules  dried  up,  and  in  about  a  fortnight  the  crusts  with  patches 
of  hair  fell  off,  leaving  more  or  less  marked  scars.  The  eruption 
appeared  at  the  same  time  on  different  parts  of  the  body,  especially 
on  the  nostrils,  lips,  buttocks,  and  vulva.  Sarrans  believed  that  the 
mares  taken  to  the  breeding  establishment  at  Rieumes  had  been 
infected  from  the  ropes  which  had  been  used  in  tying  up  other  affected 
animals,  and  had  become  thereby  infected  with  the  virus  of  this 
disease.  One  of  the  mares  was  taken  by  the  owner,  M.  Corail,  to 
the  veterinary  school  to  be  examined  by  M.  Lafosse.  About  eight 
days  after  this  visit  significant  symptoms  appeared  :  loss  of  appetite, 
lameness,  stiffness  of  both  pastern  joints,  and  a  hot,  painful  swelling 
of  the  left  pastern  joint.  The  hair  was  staring,  and  there  were 
vesicles  on  the  skin,  from  which  a  liquid  exuded  having  an  ammoniacal 
odour  but  less  foetid  than  the  secretion  in  eaux  auxjambes. 

M.  Lafosse  successfully  transmitted  the  disease  to  cows,  and  from 
cows  to  children  and  to  a  horse. 

In  1863  the  subject  of  vaccinogenic  grease  or  horse-pox  again 
received  great  attention  in  France.  A  student  named  Amyot  was 
engaged  in  dressing  a  horse  on  which  an  operation  had  been  per- 
formed. The  leg  which  had  been  operated  on  became  the  seat  of  a 
very  confluent  eruption  of  horse- pox,  which  was  followed  by  such 
an  abundant  flow  of  serosity  that  at  first  the  nature  of  the  affection 
was  mistaken,  and  it  was  thought  to  be  a  complication  of  eaux  aux 


308  INFECTIVE    DISEASES. 

jambes.  Amyot  had  a  wound  on  the  dorsal  aspect  of  the  first  inter- 
phalangeal  joint  of  the  little  finger  of  his  right  hand  ;  in  spite  of 
this,  he  continued  to  dress  the  horse  entrusted  to  his  care.  The 
wound  on  his  finger  became  accidentally  inoculated  with  the  virus, 
which  flowed  in  great  abundance  from  the  horse's  leg. 

The  wound  was  made  on  August  3rd,  and  the  next  day  it  was 
swollen,  and  rather  painful.  On  the  5th,  Amyot  suffered  from  malaise 
and  great  weakness;  on  the  6th,  7th,  and  8th,  vesicles  appeared 
successively  on  the  fingers  of  his  left  hand,  and  on  his  forehead 
between  the  two  eyebrows.  On  the  9th,  these  vesicles  were  fully 
developed ;  those  of  the  fingers  consisted  of  very  large  epidermic 
bullse  on  a  bluish-red  base.  On  opening  them,  a  perfectly  limpid 
fluid  escaped  in  such  abundance  that  small  test-tubes  might  have 
been  filled  with  it.  The  vesicle  on  the  forehead  was  surrounded  by 
a  bluish- red  areola,  within  which,  the  epidermis,  of  a  leaden-grey 
hue,  was  raised,  and  had  a  slight  central  depression.  The  liquid 
which  flowed  from  it  when  it  was  opened,  and  which  continued  to 
ooze,  was  also  very  abundant  and  of  a  deep  citrine  colour. 

The  vesicles  which  had  developed  on  the  dorsal  side  of  Amyot's 
fingers  were  extremely  painful.  The  incessant  shooting  pains,  of 
which  they  were  the  seat,  prevented  him  from  getting  any  rest  for 
three  days.  On  the  10th,  inflammation  of  the  lymphatics  followed  ; 
both  arms  were  swollen  and  very  painful,  with  red  lines  indicating 
the  course  of  the  lymphatic  vessels.  The  glands  of  the  axillae  were 
also  enlarged. 

The  lymphatic  glands  behind  the  jaws  were  also  swollen  and  pain- 
ful. Amyot's  chief  sufferings  were  occasioned  by  the  intense  local 
pain  caused  by  the  vesicles  on  the  fingers,  and  by  the  inflammation 
of  the  lymphatic  vessels  and  glands,  which  continued  in  this  state  up 
to  the  18th  of  August.  It  was  only  at  the  end  of  the  month  that  the 
vesicles  were  completely  cicatrised. 

Bouley  felt  very  great  anxiety  in  the  presence  of  the  grave 
symptoms  which  accompanied  the  eruption.  The  eruption  on  the 
forehead  was  especially  a  cause  of  great  uneasiness,  because  glanders 
manifests  itself  in  a  similar  way. 

With  virus  from  Amyot's  vesicles  the  disease  was  transmitted  to 
cows  and  to  children. 

Further,  this  outbreak  enabled  exhaustive  experiments  to  be 
made,  by  which  it  was  definitely  established  that  horse-pox  is  never 
infectious,  but,  like  cow-pox,  is  transmitted  solely  by  contact. 

In  1880,  M.  Baillet,  Director  of  the  National  Veterinary  School 
of  Toulouse,  was  informed  that  a  contagious  malady  had  developed 


CONSTITUTIONAL   GREASE   OR   HORSE-POX.  309 

in  the  mares,  which  had  been  served  by  the  stallions  at  the  breeding 
establishment  at  Rieumes,  belonging  to  M.  Mazeres.  M.  Peuch 
was  delegated  to  investigate  this  outbreak,  and  he  visited  for  that 
purpose  Berat,  Rieumes,  and  Labastide-Clermont. 

At  Berat  three  mares  were  examined.  In  one,  there  were  scars 
and  crusts,  the  remains  of  an  eruption  on  the  lips  and  in  the  vicinity 
of  the  vulva  ;  in  another,  there  were  several  reddish  circular  ulcers 
in  the  same  region ;  and  in  a  third,  there  were  dried  pustules  with 
blackish  adherent  crusts  at  the  circumference  of  the  vulva  and 
extending  over  the  perinseum.  On  the  lower  part  of  the  left  flank 
a  vesicle  was  discovered  surmounted  by  a  crust,  and  when  the  latter 
was  detached  a  sero-sanguinolerit  liquid  oozed  from  the  exposed 
surface.  M.  Peuch  recognised  the  true  nature  of  this  disease, 
having  several  times  previously  had  the  opportunity  of  examining 
mares  with  a  vesicular  eruption  round  the  vulva  after  coition, 
which  eruption  he  had  studied  from  its  first  appearance  to  complete 
cicatrisation,  and  had  ascertained  to  be  horse-pox. 

On  proceeding  to  Rieumes,  M.  Peuch  inspected  eleven  stallions, 
six  horses,  and  five  asses.  In  one  ass  there  were  several  vesicles 
on  the  right  side  of  the  penis  scattered  about  from  the  base  to  the 
glans.  In  another  ass  there  was  a  trace  of  a  vesicle  on  the  penis 
and  a  characteristic  vesicle  on  the  left  nostril. 

In  an  old  bay  mare  there  were  the  remains  of  an  eruption 
on  the  circumference  of  the  vulva,  and  in  an  old  white  mare  there 
were  not  only  vesicles  on  the  vulva,  but  in  addition  vesicles  on  the 
inner  side  of  the  lower  lip.  M.  Peuch  drew  special  attention  to 
these  cases  as  likely  to  be  confounded  with  aphthous  stomatitis,  but 
the  existence  of  the  same  eruption  on  other  parts  of  the  body  is  an 
important  aid  in  making  a  diagnosis  of  horse-pox. 

At  Labastide-Clermont  one  mare  was  particularly  noticed. 
This  mare  had  been  served  on  the  19th  and  21st  of  April,  and  on  the 
occasion  of  the  inspection,  May  llth,  there  were  the  remains  of  an 
eruption  around  the  vulva,  and  lymphangitis  existed  in  the  right 
posterior  limb,  which  was  engorged,  hot,  and  painful  in  its  whole 
extent,  so  that  the  animal  walked  with  difficulty.  The  proprietor 
had  contracted  the  disease  in  attending  to  his  mare,  and  exhibited 
a  vesicle  on  the  thumb  of  the  right  hand,  excoriated  and  blackened, 
but  still  recognisable. 

Some  of  the  crusts  collected  from  the  cases  at  Berat  were  used 
for  inoculating  a  cow.  The  result  was  successful,  and  the  disease 
was  transmitted  by  inoculation  to  a  heifer  and  several  students  and 
children. 


310  INFECTIVE    DISEASES. 

M.  Peuch  ascertained  that  horse-pox  caused  considerable  alarm 
from  the  fact  that  the  breeders  regard  this  eruptive  affection  as 
syphilitic,  and  this  alarm  consequently  brings  discredit  upon  the 
breeding  establishment  whence  the  illness  has  spread.  He  was  also 
led  to  appreciate  the  great  necessity  for  further  study  of  this  disease 
in  relation  to  dourine  or  maladie  du  coit. 

In  1882  M.  Peuch  had  the  opportunity  of  investigating  a  case 
of  horse-pox  in  Algeria.  The  disease  occurred  in  a  thoroughbred 
Arab.  There  was  an  eruption  of  vesicles,  and  there  was  also  an 
ulcer  the  size  of  a  five-franc  piece  in  the  nostril.  In  the  mouth  and 
on  the  lips  there  were  a  number  of  small  vesicles  about  the  size  of 
a  pea.  The  sublingual  glands  were  engorged,  hot,  and  painful  on 
pressure.  The  coat,  in  patches,  on  the  lateral  aspect  of  the  neck, 
on  the  shoulders,  the  flanks,  and  in  the  hollow  of  the  heel,  was 
staring,  giving  the  appearance  of  small  paint-brushes.  On  passing 
the  hand  over  these,  vesicles  could  be  detected  partly  dry  and  partly 
secreting. 

The  disease  was  transmitted  to  cows,  and  from  cows  to  about 
one  thousand  five  hundred  persons. 

Cases  similar  to  the  one  just  described,  in  which  there  is  more  or 
less  marked  ulceration  of  the  nostril  or  nasal  septum,  must  be  care- 
fully distinguished  from  glanders.  And  again,  when  the  sublingual 
glands  are  affected  the  disease  may  be  mistaken  for  strangles. 

NATURE  AND  AFFINITIES. 

Horse-pox  and  human  small-pox  are  quite  distinct  diseases, 
and  the  theory  that  horse-pox  is  derived  from  grooms  or  other 
attendants  suffering  from  small-pox  may  be  dismissed  without 
further  comment. 

Hor^e-pox  is  never  infectious,  but  is  communicated  solely  by 
contact — either  by  grooms  inoculating  the  virus  with  their  hands, 
sponges,  or  brushes,  or  by  horses  coming  into  contact  with  each  other, 
and  in  breeding  establishments  by  coition.  Auzias  Turenne,  who 
wrote  exhaustively  on  this  subject,  maintained  that  horse-pox  came 
into  the  same  category  of  diseases  as  syphilis  in  man. 

"A  un  point  de  vue,  le  grease  pustuleux  inocule  offre  la  plus 
parfaite  ressemblance  avec  la  verole  inoculee,  par  le  produit  des 
accidents  secondaires.  Des  deux  cotes  nous  voyons,  absence  de 
contagion  par  la  voie  de  1'atmosphere,  travail  local,  retentissement 
lymphatique  et  ganglionaire,  fermentation  universelle  de  Forganisme, 
eruption  gerierale  et  immunite  acquise  centre  de  nouvelles  atteintes. 

"  A  un  autre  point  de  vue,  la  ressemblance  avec  la  variole  est 


NATURE    AND   AFFINITIES.  311 

frappante.     Mais  il  s'en  distingue  eriorniement  par  1'absence  de  la 
contagiosite  atmospherique." 

Human  small-pox  belongs  to  a  different  group  of  diseases,  and 
has  affinities  rather  with  small-pox  of  sheep  and  cattle  plague, 
diseases  which  are  not  only  inoculable,  but  highly  infectious.  Human 
small-pox  is  an  infectious  disease  characterised  by  sudden  and  severe 
fever,  followed  after  forty-eight  hours  by  a  generalised  eruption ;  horse  - 
pox  commences  as  a  local  affection,  and  constitutional  symptoms 
follow.  Auzias  Turenne,  guided  by  analogy,  described  the  general- 
ised eruptions  following  "grease"  or  horse-pox  as  greasides  ("comme 
on  dit  syphilides"). 

Horse-pox  and  human  syphilis  are  absolutely  distinct  diseases; 
and  there  is  no  more  ground  for  believing  that  horse-pox  originates 
in  human  syphilis  than  there  is  for  accepting  the  theory  that  it  arises 
from  grooms  suffering  from  small-pox.  Syphilis  artificially  inocu- 
lated on  the  human  subject  only  resembles  the  casual  or  intentional 
inoculation  of  virulent  horse-pox.  The  stages  of  papulation,  vesicu- 
lation,  ulceration,  scabbing,  and  the  formation  of  a  permanent  scar, 
occur  in  inoculated  syphilis,  and  if  we  examine  Ricord's  illustrations 
and  study  the  experiments  of  Auzias  Turenne,  we  cannot  fail  to 
be  struck  with  the  remarkable  similarity  to  the  results  obtained  and 
depicted  by  Jeimer. 

But  in  order  to  follow  the  argument  of  Auzias  Turenne  we 
must  study  the  natural  and  casual  horse-pox.  And  if  we  are  not 
familiar  with  what  has  been  written  on  this  subject,  and  if  we 
restrict  our  knowledge  to  the  artificially  cultivated  horse- pox,  we 
shall  fail  to  recognise  the  disease  when  we  meet  with  it,  and  we  shall 
be  liable  to  attribute  the  results  of  the  full  effect  of  the  virus  to 
accidental  contamination. 

Another  question  of  very  great  interest  is  the  relation  of  horse- 
pox  to  cow-pox.  Jenner  first  of  all  propounded  the  theory  that  all 
cow-pox  arose  from  horse-pox,  or  as  he  termed  it  "  the  grease,"  and 
thus  cow-pox  and  horse- pox  were  manifestations  of  the  same  disease. 
But  it  was  established  that  cow-pox  also  arose  quite  independently 
of  horse-pox,  and  Jenner  was  led  to  distinguish  between  cow-pox, 
a  disease  peculiar  to  the  cow,  and  the  eruptive  affection  transmitted 
to  the  cow  from  the  horse,  which  farmers  and  others,  by  a  strange 
perversion  of  terms,  called  the  cow-pox.  Whether  the  eruption  of 
cow-pox  can  be  distinguished  from  the  eruption  of  horse-pox  com- 
municated to  the  cow,  and  whether  cow-pox  and  horse-pox  are 
identical,  or  only  analogous,  are  questions  which  call  for  further 
investigation. 


312  INFECTIVE   DISEASES. 

Bacteria  in  Horse-pox. — Outbreaks  of  horse-pox  have  not 
been  investigated  from  the  bacteriological  point  of  view,  and  the 
nature  of  the  contagium  is  unknown. 

COW-POX. 

Cow-pox  is  a  vesicular  disease  of  the  teats  of  cows.  It  is  never 
infectious,  and  only  attacks  cows  in  milk,  the  virus  being  transferred 
from  cow  to  cow  by  the  hand  of  the  milker.  The  disease  is  com- 
municable to  milkers,  and  the  virus  artificially  inoculated  produces 
what  is  commonly  known  as  vaccinia.  In  its  clinical  history  and 
epidemiology  cow-pox  is  totally  distinct  from  human  small-pox,  and 
the  hypothetical  and  entirely  erroneous  suggestion  that  the  disease 
arises  from  milkers  suffering  from  human  small-pox  is  responsible 
for  the  belief  which  prevailed  until  recently,  that  cow-pox  was  an 
extinct  disease  in  this  country ;  but,  by  the  author's  researches,  this 
has  been  shown  to  be  a  mistake. 

Cow-pox  is  not  a  rare  disease,  and  it  has  never  been  found  to 
arise  from  a  milker  suffering  from  small-pox.  As  this  is  a  matter 
of  great  importance  in  discussing  the  etiology  of  the  disease,  the 
history  of  outbreaks  and  the  clinical  characters  of  cow-pox  will  be 
given  in  considerable  detail. 

According  to  Jenner,  cow-pox  had  been  known  among  farmers 
from  time  immemorial.  He  refers  to  cases  occurring  in  1770,  1780, 
1782,  1791,  1794,  1796,  and  1798.  In  1799  cow-pox  was  raging  in 
the  dairies  in  London,  and  outbreaks  were  investigated  by  Woodville, 
Pearson,  and  Bradley.  In  the  same  year  cow-pox  broke  out  at 
Norton  Nibley,  in  Gloucestershire.  Pearson  and  Aikin  referred  to 
the  prevalence  of  cow-pox  in  Wilts,  Somerset,  Devon,  Bucks,  Dorset, 
Norfolk,  Suffolk,  Leicestershire,  and  Staffordshire  ;  and  Barry  men- 
tioned its  prevalence  in  Ireland. 

From  this  time  onwards,  for  a  long  period,  natural  cow-pox 
received  little  or  no  attention  in  this  country.  Fresh  stocks  of  lymph 
were  raised  for  the  purposes  of  vaccination,  but  no  further  attention 
was  given  to  studying  the  disease  in  the  cow.  In  1836  Leese 
described  an  outbreak  of  cow-pox,  and  in  1838  Estlin  discovered  an 
outbreak  in  Gloucestershire.  In  1838-39  cow-pox  was  met  with 
by  Mr.  Fox,  of  Cerne  Abbas,  and  again  in  1839,  in  Dorsetshire,  by 
Mr.  Sweeting.  Ceely  frequently  met  with  cow-pox  in  the  Vale  of 
Aylesbury,  and  particularly  refers  to  outbreaks  in  1838,  1840,  1841, 
and  1845.  But  after  this,  outbreaks  of  this  disease  in  the  cow  were 
not  recorded,  though  several  medical  practitioners  met  with  the 
disease  and  raised  fresh  stocks  of  vaccine  lymph.  Thus,  when 


COW-POX.  313 

inquiries  were  made  in  1857,  it  was  found  that  Mr.  Donald 
Dalrymple,  of  Norwich  (on  two  occasions),  Mr.  Beresford,  of  Nar- 
borough,  in  Leicestershire,  Mr.  Gorham,  of  Aldeburgh,  Mr.  Alison, 
of  Great  Retford,  Mr.  Coles,  of  Leckhampton,  Mr.  Rudge,  of 
Leominster,  and  one  or  two  others,  had  met  with  outbreaks  of 
cow-pox. 

In  1885  cow-pox  was  discovered  by  the  author  in  Wiltshire.  The 
publication  of  the  fact  led  to  the  recognition  of  the  disease  in  the 
same  year  in  many  parts  of  England,  and  cases  were  met  with  in 
man  in  1888  by  Mr.  Forty  in  Gloucestershire,  and  by  Mr.  Bucknill 
near  London  in  1894. 

In  Italy,  cow-pox  was  found  by  Sacco  in  the  plains  of  Lombardy 
in  1800,  and  by  other  practitioners  in  1808-9.  In  1812  it  was 
observed  at  Naples  by  Miglietta;  in  1830  in  Piedmont ;  and  in  1832 
and  1843  at  Rome,  by  Dr.  Maceroni.  More  recently,  several  out- 
breaks of  cow-pox  have  been  met  with  in  this  country,  and  the  stocks 
of  vaccine  lymph  renewed. 

In  France,  in  1810,  cow-pox  was  found  in  the  department  of 
La  Meurthe,  and  in  1822  at  Clairvaux ;  at  Passy,  Amiens,  and 
Rambouillet  in  1836  ;  at  Rouen  in  1839  ;  at  St.  Illide,  at  St.  Seine, 
and  at  Perylhac,  in  1841  ;  in  1842  at  Pagnac;  in  1843  at  Deux 
Jumeaux,  where,  during  the  previous  thirty  years,  several  fresh 
stocks  of  lymph  had  been  raised  and  circulated.  The  disease  occurred 
in  a  cow  belonging  to  M.  Majendie  in  1844,  and  it  was  found  at 
Wasseloune,  in  the  department  of  Bas  Rhin,  in  1845  ;  it  occurred  in 
three  other  departments  in  1846  ;  at  Rheims,  and  in  the  department 
of  Eure  et  Loire,  in  1852 ;  in  the  arrondissement  of  Sancerre,  and  at 
Beziers  in  1854;  and  at  Guyonville  in  1863.  It  broke  out  on  farms 
in  three  villages  near  Nogent  in  1864  (the  disease  was  introduced  by 
newly  purchased  cows  ;  milkers  were  infected,  and  from  one  of  these 
milkers  a  lymph  stock  was  established);  it  also  occurred  in  1864,  at 
Petit  Quevilly,  near  Rouen;  and  in  April  1866  at  Beaugency ;  in 
1881  at  Eysines,  near  Bordeaux,  and  again  at  the  same  place  in 
1883  ;  and  in  1844  at  Cerons. 

In  Germany,  as  soon  as  attention  had  been  drawn  to  the  disease, 
cow-pox  was  frequently  discovered.  There  were  as  many  as  thirty- 
eight  outbreaks  reported  in  one  year  in  Wurtemberg. 

It  is  hardly  necessary,  after  reciting  these  instances,  to  insist 
that  cow-pox  is  far  from  being  a  rare  disease,  as  many  have  sup- 
posed who  are  unacquainted  with  the  literature  of  the  subject  and 
unfamiliar  with  the  appearances  of  the  natural  disease  in  the 
cow. 


314  INFECTIVE    DISEASES. 

NATURAL  AND  CASUAL  COW-POX. 

To  appreciate  the  characters  of  the  natural  disease  in  the  cowy 
we  must  dismiss  from  our  minds  the  artificial  disease  vaccinia,  for 
the  ordinary  results  of  vaccination  stand  in  much  the  same  relation 
to  the  natural  disease  cow-pox  as  the  benign  vesicle  of  variolation  to 
natural  small-pox. 

The  description  of  cow-pox  given  by  Jenner,  in  1798,  was  the 
first  published  account.  The  disease  in  the  cow  was  described  as 
consisting  of  irregular  pustules  on  the  teats,  of  a  palish  blue  colour, 
surrounded  by  an  erysipelatous  inflammation,  and  characterised  by 
a  tendency  to  degenerate  into  phagedsenic  ulcers.  The  animals  were 
indisposed  and  the  secretion  of  milk  lessened. 

In  referring  to  an  outbreak  which  occurred  epizootically  in 
London  in  February  1799,  Dr.  Bradley  gave  a  coloured  plate  of  the 
disease  on  the  arm  and  fingers  of  a  milker.  The  cow-pox,  he  said, 
in  this  instance,  "  appears  to  have  been  very  mild,  for  no  loss  was 
experienced  by  the  farmers  from  the  deficiency  of  milk,  as  usually 
happens." 

These  early  descriptions  were  supplemented  by  an  account  of 
cow'-pox  by  Mr.  Lawrence,  author  of  A  Philosophical  and  Practical 
Treatise  on  Horses,  and  on  the  Moral  Duties  of  Man  toward  the  Brute 
Creation.  Lawrence's  article  on  cow-pox  not  only  affords  evidence 
that  this  disease  was  known  to  those  who  had  the  care  of  cattle 
before  Jenner's  paper  was  published,  but  it  shows  that  it  had  also 
been  made  the  subject  of  practical  observation  and  study  by  veteri- 
narians. Lawrence  concluded  by  saying  :  "  Whatever  may  be  the 
fate  of  cow-pox  inoculation,  it  has  and  will  give  further  occasion  to 
a  pretty  large  and  open  discussion,  which  is  always  beneficial  as 
having  a  tendency  to  produce  discovery  and  promote  improvement ; 
and  when  the  public  ardour  for  the  present  topic  shall  have  become 
a  little  cool  and  satisfied,  I  hope  it  will  be  turned  by  enlightened 
men  towards  another,  perhaps  of  nearly  as  great  consequence — 
namely,  the  prevention  of  the  original  malady  in  the  animals  them- 
selves. Those  who  have  witnessed  it  and  only  reflected  upon  the 
excessive  filth  and  nastiness  which  must  unavoidably  mix  with  the 
milk  in  an  infected  dairy  of  cows,  and  the  corrupt  and  unsalubrious 
state  of  their  produce  in  consequence,  will  surely  join  me  in  that 
sentiment." 

Lawrence  was  almost  a  century  before  his  time.  Cow-pox  was  not 
again  brought  forward  in  this  light  until  1887-88,  when  the  author 
reported  the  contamination  of  the  milk  at  the  Wiltshire  farms,  and 


NATURAL   AND   CASUAL   COW-POX.  315 

advocated  the  advisability  of  placing  this  disease  under  the  Con- 
tagious Diseases  (Animals)  Act. 

The  numerous  pathological  details  wanting  in  the  early  accounts 
of  cow-pox  were  supplied  by  the  painstaking  and  laborious  re- 
searches of  Robert  Oeely.  From  his  classical  papers  in  the  Trans- 
actions of  the  Provincial  Medical  Association,  we  can  obtain  a 
complete  picture  of  the  natural  disease  in  the  cow. 

In  Ceely's  experience  in  the  Yale  of  Aylesbury,  outbreaks 
occurred  at  irregular  intervals,  most  commonly  appearing  about 
the  beginning  or  end  of  the  spring,  rarely  during  the  height  of 
summer.  There  were  outbreaks  at  all  periods  from  August  to  May 
and  the  beginning  of  June,  cases  being  met  with  in  autumn  and 
the  middle  of  winter,  after  a  dry  summer.  The  disease  was  occa- 
sionally epizootic,  or  occurring  at  times  at  several  farms  at  no  great 
distance  from  each  other,  but  was  more  commonly  sporadic  or 
nearly  solitary.  It  was  to  be  seen  sometimes  at  several  contiguous 
farms  ;  at  other  times  at  one  or  two  farms.  Many  years  might 
elapse  before  it  recurred  at  a  given  farm,  although  all  the  animals 
might  have  been  changed  in  the  meantime.  Cow-pox  had  broken 
out  twice  in  five  years  in  a  particular  vicinity  at  two  contiguous 
farms,  while  at  an  adjoining  dairy,  in  all  respects  similar  in  local 
and  other  circumstances,  it  had  not  been  known  to  exist  for  forty 
years.  It  was  sometimes  introduced  into  a  dairy  by  recently 
purchased  cows.  Twice  it  had  been  known  to  be  so  introduced  by 
milch  heifers.  It  was  considered  that  the  disease  was  peculiar  to 
the  milch  cow  ;  it  came  primarily  while  the  animal  was  in  milk, 
and  it  was  casually  propagated  to  others  by  the  hands  of  the  milkers. 
Sturks,  dry  heifers,  dry  cows,  and  milch  cows  milked  by  other  hands, 
grazing  in  the  same  pastures,  feeding  in  the  same  sheds,  and  at 
contiguous  stalls,  remained  exempt  from  the  disease. 

For  many  years,  the  "spontaneous"  origin  of  cow-pox  had 
not  been  doubted  in  the  Yale  of  Aylesbury.  In  all  the  cases  that 
Ceely  had  noticed  he  could  never  discover  the  probability  of  any 
other  origin. 

Condition  of  Animal  primarily  affected. — There  was  much  diffi- 
culty in  determining  at  all  times,  with  precision,  whether  this 
disease  arose  primarily  in  one  or  more  individuals  in  the  same  dairy. 
Most  commonly,  however,  it  appeared  to  be  solitary.  The  milkers 
believed  they  were  able  to  point  out  the  infecting  individual.  In 
two  instances,  there  could  be  very  little  doubt  on  this  point.  In 
August  1838,  three  cows  were  affected  with  the  disease.  The  first 
was  attacked  two  months  after  calving  and  seven  weeks  after 


316  INFECTIVE    DISEASES. 


weaning.  This  animal  was  considered  in  good  health,  but  it  looked 
out  of  condition.  Heat  and  tenderness  of  the  teats  and  udder  were 
the  first  noticed  signs.  The  other  two  were  affected  in  about  ten 
days.  In  December  1838,  in  a  large  dairy,  a  milch  cow  slipped  her 
calf,  had  heat  and  induration  of  the  udder  and  teats,  with  cow-pox 
eruption,  and  subsequently  leucorrhcea  and  greatly  impaired  health ; 
the  whole  dairy,  consisting  of  forty  cows,  became  subsequently 
affected,  and  also  some  of  the  milkers.  In  another  dairy,  at  the 
.same  time,  it  first  appeared  in  a  heifer  soon  after  weaning,  and  in 
about  ten  or  twelve  days  extended  to  five  other  heifers  and  one  cow, 
milked  in  the  same  shed,  affecting  the  milkers.  And  in  another 
dairy  thirty  cows  were  severely  affected,  and  also  one  of  the  milkers. 
It  appeared  to  originate  in  a  cow  two  months  after  calving.  The 
only  symptoms  noticed  were  that  the  udder  and  teats  were  tumid, 
tender,  and  hot  just  before  the  disease  appeared. 

Condition  of  Animals  casually  affected. — In  some  animals,  it  was 
less  severe  than  in  others,  depending  on  the  state  and  condition  of 
the  skin  of  the  parts  affected,  and  the  constitution  and  habit  of  the 
animal.  It  was  sometimes  observed  to  dimmish  the  secretion  of 
milk,  and  in  most  cases  it  commonly  did  actually  affect  the  amount 
artificially  obtained  ;  with  this  exception,  and  the  temporary  trouble, 
and  accidents  to  the  milk  and  the  milkers,  little  else  was  observed ; 
the  animal  continued  to  feed  and  graze  apparently  as  well  as  before. 
The  topical  effects  varied  very  much  in  different  individuals  ;  the 
mildness  or  severity  being  greatly  influenced  by  temperament  and 
condition  of  the  animal,  and  especially  by  the  state  of  the  teats  and 
udder,  and  the  texture  and  vascularity  of  the  skin  of  the  parts 
affected.  Where  the  udder  was  short,  compact,  and  hairy,  and  the 
skin  of  the  teats  thick,  smooth,  tense,  and  entire,  or  scarcely  at  all 
chapped,  cracked,  or  fissured,  the  animal  often  escaped  with  a  mild 
affection,  sometimes  with  only  a  single  vesicle.  But  where  the 
udder  was  voluminous,  flabby,  pendulous,  and  naked,  the  teats 
long  and  loose,  and  the  skin  corrugated,  thin,  fissured,  rough,  and 
unequal,  then  the  animal  scarcely  ever  escaped  a  copious  eruption. 
Hence,  in  general,  heifers  suffered  least,  and  cows  most,  from  the 
milkers'  manipulations. 

Progress  of  the  Disease. — Cow-pox  once  arising  or  introduced,  and 
the  necessary  precautions  not  being  adopted  in  time,  appeared  in  ten 
or  twelve  days  on  many  more  cows  in  succession,  so  that  among 
twenty-five  cows  perhaps  by  the  third  week  nearly  all  would  be 
affected  ;  but  five  or  six  weeks  or  more  were  required  before  the 
teats  were  perfectly  free  from  the  disease. 


NATURAL   AND   CASUAL   COW-POX.  317 

Propagation  by  the  Hand  of  the  Milker. — Ceely  was  able  to  confirm 
the  way  in  which  the  disease  was  said  to  spread.  In  December 
1838,  on  a  large  dairy  farm,  w^here  there  were  three  milking-shedsr 
cow-pox  broke  out  in  the  home  or  lower  shed.  The  cows  in  this 
shed  being  troublesome,  the  milker  from  the  upper  shed,  after 
milking  his  own  cows,  came  to  assist  in  this  for  several  days,  morning 
and  evening,  when  in  about  a  week  some  of  his  own  cows  began  to 
exhibit  the  disease.  It  appears  that,  having  chapped  hands,  he 
neglected  washing  them  for  three  or  four  days  at  a  time,  and  thus 
conveyed  the  disease  from  one  shed  to  another.  During  the  progress 
of  the  disease  through  this  shed,  one  of  the  aft'ected  cows,  which  had 
been  attacked  by  the  others,  was  removed  to  the  middle  shed,  where 
all  the  animals  were  perfectly  well.  This  cow,  being  in  an  advanced 
stage  of  the  disease,  and  of  course  difficult  to  milk  and  dangerous 
to  the  milk- pail,  was  milked  first  in  order  by  the  juvenile  milker  for 
three  or  four  days  only,  when,  becoming  unmanageable  by  him,  its 
former  milker  was  called  in  to  attend  exclusively  to  it.  In  less  than 
a  week,  all  the  animals  of  this  shed  showed  symptoms  of  the  disease, 
though  in  a  much  milder  degree  than  it  had  appeared  in  the  other 
sheds,  fewer  manipulations  having  been  performed  by  an  infected 
hand. 

Topical  Symptoms  of  the  Natural  Disease. — For  these,  Ceely  was- 
almost  always,  in  the  early  stage,  compelled  to  depend  on  the  obser- 
vations and  statements  of  the  milkers.  They  stated  that  for  three 
or  four  days,  without  any  apparent  indisposition,  they  noticed  heat 
and  tenderness  of  the  teats  and  udder,  followed  by  irregularity  and 
pimply  hardness  of  these  parts,  especially  about  the  bases  of  the 
teats  and  adjoining  the  vicinity  of  the  udder ;  these  pimples  on  skins 
not  very  dark  are  of  a  red  colour,  and  generally  as  large  as  a  vetch 
or  a  pea,  and  quite  hard,  though  in  three  or  four  days  many  of 
these  increase  to  the  size  of  a  horse-bean.  Milking  is  genera  lly 
very  painful  to  the  animal ;  the  tumours  rapidly  increase  in  size, 
vesicate,  and  are  soon  broken  by  the  hands  of  the  milker.  Milking 
now  becomes  a  troublesome  and  occasionally  a  dangerous  process 
Ceely  adds:  "It  is  very  seldom  that  any  person  competent  to 
judge  of  the  nature  of  the  ailment  has  access  to  the  animal  before 
the  appearance  of  the  disease  on  others  of  the  herd,  when  the  cow 
first  affected  presents  on  the  teats  acuminated,  ovoid,  or  globular 
vesications,  some  entire,  others  broken,  not  infrequently  two  or  three 
interfluent ;  those  broken  have  evidently  a  central  depression  with 
marginal  induration  ;  those  entire,  being  punctured,  diffuse  a  more 
or  less  viscid  amber-coloured  fluid,  collapse,  and  at  once  indicate  the 


318  INFECTIVE    DISEASES. 

same  kind  of  central  and  marginal  character.  They  appear  of 
various  sizes,  from  that  of  a  pin's  head,  evidently  of  a  later  date, 
either  acuminated  or  depressed,  to  that  of  an  almond  or  a  filbert,  or 
ever  larger.  Dark  brown,  or  black,  solid,  uniform  crusts,  especially 
on  the  udder  near  the  base  of  the  teats,  are  visible ;  at  the  same  time, 
some  much  larger  are  observed  on  the  teats ;  these,  however,  are 
less  regular  in  form  and  less  perfect.  Some  are  nearly  detached, 
others  quite  removed,  exhibiting  a  raw  surface  with  a  slight  central 
slough.  On  the  teats,  the  crusts  are  circular,  oval,  oblong,  or 
irregular  ;  some  flat,  others  elevated,  some  thin  and  more  trans- 
lucent, being  obviously  secondary.  The  appearance  of  the  disease 
in  different  stages,  or  at  least  the  formation  of  a  few  vesicles  at 
different  periods,  seems  very  evident.  The  swollen,  raw,  and  en- 
crusted teats  seem  to  produce  uneasiness  to  the  animal  only  while 
subjected  to  the  tractions  of  the  milkers,  which  it  would  appear  are 
often  nearly  as  effectual  as  usual."  Referring  again  to  the  character 
of  the  vesicle,  Ceely  says,  that  "  those  fortunate  enough  to  have  an 
opportunity  of  watching  the  disease  in  its  progress  may  observe 
that,  when  closely  examined,  they  present  the  following  characters  : 
In  animals  of  dark  skin,  at  this  period,  the  finger  detects  the 
intumescent  indurations  often  better  than  the  eye,  but  when  closely 
examined  the  tumours  present  at  their  margins  and  towards  their 
centres  a  glistening  metallic  lustre  or  leaden  hue  ;  but  this  is  not 
always  the  case,  for  occasionally  they  exhibit  a  yellowish  or  yellowish- 
white  appearance." 

In  describing  the  crusts  in  detail,  Ceely  says  that  "  large  black 
solid  crusts,  often  more  than  an  inch  or  two  in  length,  are  to  be 
seen  in  different  parts  of  these  organs,  some  firmly  adherent  to  a 
raw  elevated  base,  others  partially  detached  from  a  raw,  red,  and 
bleeding  surface ;  many  denuded,  florid,  red,  ulcerated  surfaces,  with 
small  central  sloughs  secreting  pus  and  exuding  blood,  the  teats 
exceedingly  tender,  hot,  and  swollen.  ...  In  some  animals,  under 
some  circumstances,  this  state  continues  little  altered  till  the  third 
or  fourth  week,  rendering  the  process  of  milking  painful  to  the 
animal,  and  difficult  arid  dangerous  to  the  milker." 

"  In  many,  however,  little  uneasiness  seems  to  exist.  The  parts 
gradually  heal ;  the  crusts,  although  often  partially  or  entirely 
renewed,  ultimately  separate,  leaving  apparently  but  few  deep 
irregular  cicatrices,  some  communicating  with  the  tubuli  lactiferi, 
the  greater  part  being  regular,  smoothly  depressed,  circular,  or  oval." 

Ceely  illustrated  his  classical  memoir  with  a  series  of  valuable 
coloured  drawings.  One  plate  is  a  faithful  picture  of  the  disease  on 


NATURAL   AND   CASUAL   COW-POX.  319 

the  teats  as  it  is  ordinarily  met  with ;  the  other  is  a  composite  picture, 
consisting  of  the  disease  as  ordinarily  observed  in  the  cow,  to  which 
is  superadded  a  number  of  depressed  vesicles  as  they  occur  in  inocu- 
lated cow-pox.  It  is,  however,  an  improvement  on  a  plate  published 
by  Sacco.  The  latter  is  an  elaborate  drawing,  representing  the  udder 
and  teats  of  a  cow,  with  an  eruption  purporting  to  be  the  natural 
cow-pox.  Jenner  had  described  a  bluish  tint  in  the  vesicles  in 
natural  cow-pox,  arid  Sacco  deliberately  represents  the  natural  disease 
by  a  highly  coloured  diagrammatic  illustration  in  which  he  depicts 
clusters  of  vesicles  of  inoculated  cow-pox,  coloured  blue,  and  with 
a  silvery  lustre. 

Hering  has  given  a  coloured  plate  of  the  natural  cow-pox.  On 
the  teats  are  a  number  of  oval  and  circular  bullous  vesicles  and 
crusts.  More  recently,  Layet  has  pointed  out  the  same  characters 
in  the  cow-pox  discovered  near  Bordeaux  in  1883  and  1884.  The 
classical  characters  of  the  inoculated  disease  were  wanting,  particu- 
larly the  central  depression.  In  Wiltshire,  the  author  could  only 
distinguish,  on  the  cow's  teats,  globular  and  broken  vesicles  and 
thick  prominent  crusts  and  ulcers,  appearances  which  had  very 
little  in  common  with  the  ordinary  results  of  vaccination. 

The  early  accounts  of  the  severe  character  of  the  disease  will 
appear  by  no  means  exaggerated  to  those  who  have  had  an  oppor- 
tunity of  studying  the  effects  on  the  hands  of  the  milkers,  or  indeed 
to  those  who  have  made  themselves  familiar  with  the  descriptions 
given  by  Jenner,  in  some  of  his  cases  :  — 

"  Joseph  Merret  had  several  sores  on  his  hands,  swelling  and  stiffness 
in  each  axilla,  and  much  indisposition  for  several  days. 

"  Mrs.  H.  had  sores  upon  her  hands  which  were  communicated  to  her 
nose,  which  became  inflamed  and  very  much  swollen. 

"  Sarah  Wynne  had  cow-pox  in  such  a  violent  degree  that  she  was 
confined  to  her  bed,  and  unable  to  do  any  work  for  ten  days. 

"  William  Rod  way  was  so  affected  by  the  severity  of  the  disease  that 
he  was  confined  to  his  bed. 

"  William  Smith  had  several  ulcerated  sores  on  his  hands,  and  the 
usual  constitutional  symptoms,  and  was  affected  equally  severely  a  second 
and  a  third  time. 

"  Wrlliam  Stinchcomb  had  his  hand  very  severely  affected  with  several 
corroding  ulcers,  and  a  considerable  tumour  in  the  axilla. 

"  Sarah  Nelmes  had  a  large  pustulous  sore  on  the  hand,  and  the  usual 
symptoms. 

"A  girl  had  an  ulceration  on  the  lip  from  frequently  holding  her 
finger  to  her  mouth  to  cool  the  raging  of  a  cow-pox  sore  by  blowing 
upon  it. 


320  INFECTIVE    DISEASES. 

"  A  young  woman  had  cow-pox  to  a  great  extent,  several  sores  which 
maturated  having  appeared  on  the  hands  and  wrists. 

"  A  young  woman  had  several  large  suppurations  from  cow-pox  on  the 
hands." 

Pearson  in  his  investigations  encountered,  and  was  informed  of t 
similar  experiences. 

"  Thomas  Edinburgh  was  so  lame  from  the  eruption  of  cow-pox  on 
the  palm  of  the  hand  as  to  necessitate  his  being  for  some  time  in  hospital. 
For  three  days  he  had  suffered  from  pain  in  the  armpits,  which  were 
swollen  and  sore  to  the  touch.  He  described  the  disease  as  uncommonly 
painful,  and  of  long  continuance. 

"  A  servant  at  a  farm  informed  Pearson  that  in  Wiltshire  and 
Gloucestershire  the  milkers  were  sometimes  so  ill  as  to  lie  in  bed  for 
several  days. 

"  Mr.  Francis  said  that  cow-pox  was  very  apt  to  produce  painful  sores 
on  the  hands  of  milkers. 

"  A  servant  of  Mr.  Francis  said  that  cow-pox  affected  the  hands  and 
arms  of  the  milkers  with  painful  sores  as  large  as  a  sixpence. 

"  Mr.  Dolling  describes  the  disease  as  '  a  swelling  under  the  arm,  chilly 
fits,  etc.,  not  different  from  the  breeding  of  the  small-pox.  After  the 
usual  time  of  sickening,  namely,  two  or  three  days,  there  is  a  large  ulcer, 
not  unlike  a  carbuncle,  which  discharges  matter.' 

"  Dr.  Pulteney  described  the  disease  as  causing  'a  soreness  and  swell- 
ing of  the  axillary  glands,  as  under  inoculation  for  the  small-pox,  then 
chilliness  and  rigors  and  fevers,  as  in  the  small-pox.  Two  or  three  days- 
afterwards  abscesses,  not  unlike  carbuncles,  appear  generally  on  the  hands 
and  arms,  which  ulcerate  and  discharge  much  matter.' 

"  Mr.  Bird  wrote  a  short  account :  '  It  appears  with  red  spots  on  the 
hands,  which  enlarge,  become  roundish,  and  suppurate,  tumours  take 
place  in  the  armpit,  the  pulse  grows  quick,  the  head  aches,  pains  are  felt 
in  the  back  and  limbs,  with  sometimes  vomiting  and  delirium.' 

"  Annie  Francis  had  pustules  on  her  hands  from  milking  cows.  These 
pustules  soon  became  scabs,  which,  falling  off,  discovered  ulcerating  and 
very  painful  sores,  which  were  long  in  healing.  Some  milk  from  one  of 
the  diseased  cows,  having  spurted  on  the  cheek  of  her  sister  and  on  the 
breast  of  her  mistress,  produced  on  these  parts  of  both  persons  pustules 
and  sores  similar  to  her  own  on  her  hands." 

In  more  recent  times  these  descriptions  have  been  confirmed. 

In  1836  cow-pox  was  discovered  at  Passy,  near  Paris.  A  black 
cow,  in  very  poor  condition,  had  cow-pox  six  weeks  after  calving. 
Bousquet  had  no  opportunity  of  seeing  the  eruption  in  the  early 
stage,  but  on  examination  he  found  reddish-brown  crusts  on  the 
teats,  which  later  gave  place  to  puckered  scars.  The  milk-woman, 
Fleury,  who  had  had  small-pox,  nevertheless  contracted  the  disease 
from  the  cow.  She  had  several  vesico -pustules  on  the  right  hand 


NATURAL   AND   CASUAL   COW-POX.  321 

and  on  her  lips.  A  vesico-pustule,  when  opened  with  a  lancet, 
discharged  like  an  abscess. 

In  a  letter  to  Mr.  Badcock,  dated  April  3rd,  1845,  Ceely  referred 
to  another  new  stock  of  lymph  raised  from  a  milker's  hand.  He 
added : — 

"  In  the  enclosed  lymph  I  see  nothing  unusually  severe,  except 
on  very  thin  skins ;  although  the  milker's  hand  exhibits  now  rough 
ulcers,  one  on  the  hand  deep  enough  to  encase  a  bean." 

Recent  discoveries  of  cow-pox  in  England. — After  Ceely 's  cases  in 
1840-41,  no  cases  of  casual  cow-pox  on  the  hands  of  milkers  were 
recognised  as  such  and  recorded  in  this  country  for  nearly  fifty  years. 
In  the  outbreak  of  cow-pox  discovered  by  the  author  in  December 
1887,  in  Wiltshire,  the  disease  was  communicated  to  nearly  all  the 
milkers.  The  reader  is  referred  to  the  account  of  this  outbreak, 
which  has  already  been  given  in  the  chapter  on  scarlet  fever  (p.  274). 

The  author's  researches  were  confirmed  by  Mr.  Forty  in  1888, 
and  Mr.  Bucknill  in  1895. 

In  June  1888  Mr.  Forty,  in  practice  at  Wotton-under-Edge, 
Gloucestershire,  reported  to  the  Local  Government  Board,  that  at 
a  farm  at  Alderley,  an  eruptive  disease  on  the  udder  and  teats  was 
occurring  amongst  cows,  and  that  the  farmer's  son,  and  other  persons 
engaged  as  milkers,  had  contracted  an  eruption  like  that  of  the  cows. 
The  farmer's  son  had  been  under  Mr.  Forty's  care  suffering  from  an 
eruption,  and  circum-anal  piles.  Mr.  Forty  had  watched  the  course  of 
the  eruption  from  papules  to  vesicles  and  scabbing,  and  concluded 
that  the  eruption  could  not  be  distinguished  from  vaccinia.  Klein 
visited  the  farm,  and  found  a  number  of  cows  with  sores  on  the  teats 
and  udders.  The  sores  were  of  various  sizes  and  outline,  mostly 
irregular,  and  covered  with  brown  or  black  scabs.  Those  on  the 
teats  were  larger  and  more  irregular  than  those  on  the  udder. 
Klein  was  shown  several  milkers  who  had  had  sores  on  one  or  more 
fingers ;  one  had  had  a  bad  arm  with  swollen  axillary  glands. 
The  farmer  had  also  contracted  the  eruption ;  but  in  these  persons 
only  scabs  were  visible  as  the  remnants  of  their  sores. 

A  girl  of  about  twenty  had  taken  the  place  of  an  incapacitated 
milker,  and  noticed  a  red  pimple  form  on  the  dorsal  surface  of  her 
right  thumb.  Eight  days  afterwards  there  was  a  slightly  raised 
circular  vesicle,  with  dark  centre  and  pale  periphery ;  the  centre  of  the 
vesicle  was  slightly  depressed.  It  was  just  under  half  an  inch  in 
diameter  ;  there  was  peripheral  redness,  but  no  marked  areola.  The 
girl  had  three  good  vaccination  marks. 

Klein  experimented  on  calves  with  lymph  from  the  vesicle  and 


322  INFECTIVE   DISEASES. 

crusts  from  the  cow's  teats,  with  the  result  that  from  both  sources 
an  eruption  was  produced,  which  in  appearance  and  course  was  like 
vaccinia.  With  lymph  from  one  of  the  calves,  a  public  vaccinator 
inoculated  a  number  of  infants,  and  fine  vesicles  developed,  indis- 
tinguishable from  vaccinia. 

In  1894  Mr.  Bucknill  met  with  a  case  in  a  milkman.  He  had 
been  milking  a  cow  affected  with  cow-pox,  and  on  the  ninth  day  after 
exposure  to  infection,  and  the  seventh  day  after  the  eruption  of  the 
first  papule,  there  were  three  pocks  on  the  fore-arm.  The  pocks 
were  elevated,  circular,  and  umbilicated,  with  a  dull,  creamy-white 
ring  at  the  circumference,  and  there  was  well-marked  induration 
and  extensive  areola.  There  were  four  excellent  marks  of  primary 
vaccination.  The  vesicles  contained  clear  lymph,  and  re-inoculation 
of  the  arm  failed  to  take.  An  attempt  to  re-vaccinate  the  man 
with  current  calf  lymph  produced  only  topical  irritation. 

INOCULATED  COW-POX. 

Natural  or  Virulent  Lymph. — Severe  symptoms  are  not  limited  to 
milkers  casually  infected  from  the  cow.  Under  certain  conditions, 
artificial  inoculation  of  fresh  virus  from  the  cow  reproduces  the 
•disease  without  any  mitigation.  Thus,  in  Jenner's  cases  : — 

"  James  Phipps.  The  incisions  assumed  at  their  edges  rather  a 
darker  hue  than  in  variolous  inoculation,  and  the  efflorescence  around 
them  took  on  more  of  an  erysipelatous  look.  They  terminated  in 
scabs  and  subsequent  eschars. 

"  Susan  Phipps  was  inoculated  from  the  cow  by  inserting  matter 
into  a  superficial  scratch  on  December  2nd.  The  child's  arm  now 
showed  a  disposition  to  scab,  and  remained  nearly  stationary  for 
two  or  three  days,  when  it  began  to  run  into  an  ulcerous  state, 
and  then  commenced  a  febrile  indisposition,  accompanied  with  an 
increase  of  auxiliary  tumour.  The  ulcer  continued  spreading  near 
a  week,  during  which  the  child  continued  ill,  when  it  increased  to 
a  size  nearly  as  large  as  a  shilling.  It  began  now  to  discharge  pus  ; 
granulations  sprung  up,  and  it  healed." 

Jenner's  lymph  was  employed  by  Mr.  Cline  with  similar  results. 

"  The  child  sickened  on  the  seventh  day,  and  the  fever,  which 
was  moderate,  subsided  on  the  eleventh.  .  .  .  The  ulcer  was  not 
large  enough  to  contain  a  pea." 

Precisely  similar  experiences  have  since  been  encountered,  in  the 
early  removes  of  fresh  stocks  of  virulent  lymph.  Bousquet  in  France, 
in  his  first  trials  with  a  new  lymph,  in  1836,  made  three  punctures, 
but  he  had  soon  to  abandon  this  practice,  because  the  intensity 


INOCULATED   COW-POX.  323 

of  the  inflammation  was  sometimes  so  great  that  it  spread  over  the 
entire  arm  as  far  as  the  glands  of  the  axilla.  In  one  case,  the 
vesicles  were  enormous,  and  the  inflammation  so  violent,  that  baths, 
poultices,  fomentations,  and  antiphlogistic  diet  scarcely  sufficed 
to  reduce  it.  The  crusts  when  they  fell  off  left  ulcerations  which 
were  very  slow  to  undergo  cicatrisation.  In  some  cases,  the  vesicles 
which  resulted  hollowed  out  the  skin  so  deeply  that  they  left  regular 
holes. 

In  the  following  year  Estlin,  in  England,  started  a  stock  of  fresh 
vaccine  virus  from  the  cow,  and  found  on  inoculating  children  that 
the  new  lymph  was  extremely  active. 

In  52  the  disease  was  regular, 
,,     1  severe  erysipelas, 

,,     4  erythematous  eruptions  of  a  violent  character, 
,,     2  highly  inflamed  ulcerated  arms, 
,,     1  no  effect  after  twice  vaccinating, 
„     8  result  unknown  ;  supposed  to  have  been  favourable. 

68 

Cultivated  or  Attenuated  Lymph. — When  cow-pox  lymph  has 
been  mitigated  by  successive  transmission  through  the  human  subject, 
or  by  cultivation  on  the  belly  of  the  calf,  with  careful  selection 
of  vesicles,  it  will  produce  effects  which  are  as  follows :  About 
the  end  of  the  second  day  after  insertion,  or  early  on  the  third  day, 
a  slight  papular  elevation  is  noticeable.  By  the  fifth  or  sixth  day, 
it  has  become  a  distinct  vesicle,  of  a  bluish-white  colour,  with  raised 
margin  and  central  cup- like  depression.  By  the  eighth  day,  the 
vesicle  is  perfect.  It  is  circular,  pearl-coloured,  distended  with  clear 
lymph,  and  the  central  depression  is  well  marked.  On  the  same  day, 
or  a  little  earlier,  the  areola  begins  to  appear,  and  gradually  extends 
to  a  diameter  of  from  one  to  three  inches,  accompanied  with 
induration  and  tumefaction  of  the  subjacent  connective  tissue.  After 
the  tenth  day,  the  areola  begins  to  fade,  and  the  vesicle  at  the  same 
time  begins  to  dry  in  the  centre ;  the  lymph  becomes  opaque  and 
gradually  concretes,  and  by  the  fourteenth  or  fifteenth  day,  a  hard 
mahogany-coloured  scab  is  formed  which  contracts,  dries,  blackens,  and 
falls  off  between  the  twentieth  and  twenty-fifth  days.  A  circular, 
depressed,  foveated,  and  sometimes  radiated  scar  remains  behind. 
By  selecting  characteristic  vesicles  on  the  calf  or  on  the  human 
subject,  and  by  collecting  the  lymph  at  an  early  stage  on  the  fifth, 
sixth,  or  seventh  day,  this  artificial  disease,  commonly  known  MN 


324  INFECTIVE    DISEASES. 

vaccinia,  can  be  kept  up  in  this  comparatively  mild  form.  But 
under  certain  conditions,  such  as  a  peculiarity  in  the  subject  inocu- 
lated, or  if  lymph  be  taken  too  late,  there  will  be,  just  as  in  variolatipn, 
tendency  to  revert  to  the  full  intensity  of  the  natural  virus. 

Bacteria  in  Vaccine  Lymph. — Cohn,  Sanderson,  and  Godlee 
described  micrococci  in  vaccinal  vesicles.  Quist.  and  Ferre  in  1883 
investigated  the  same  subject.  Yoigt  in  1885  distinguished  three 
species  of  micrococcus — a  diplococcus,  a  large  coccus,  and  a  third  form. 
Bauer  in  the  same  year  described  the  presence  of  bacilli  and  sphsero- 
cocci.  Marotta  in  1886  regarded  a  tetracoccus  as  the  specific  micro- 
organism, and  Tenhot  in  1887  distinguished  a  dozen  micrococci,  two 
bacilli,  and  two  yeasts.  In  the  same  year  Garre  isolated  a  micrococcus 
which  appeared  to  him  to  be  the  contagium,  but  inoculated  on  a  child 
it  neither  produced  local  vesicles  nor  immunity  ;  while  Guttmann 
pointed  out  three  micro-organisms  which  appeared  to  be  rather  more 
constantly  present  than  others.  Pf eiffer  much  more  fully  investigated 
the  bacteriology  of  vaccine  lymph,  and  found  Saccharomyces  vaccinse, 
which  was  seldom  present  in  human  lymph  but  constantly  found  in 
calf  lymph  ;  sarcinae,  both  in  human  and  calf  lymph,  including  Sarcina 
lutea,  Sarcina  tetragonus,  Sarcina  aurantiaca,  Sarcina  muscopus ; 
bacteria  and  bacilli  were  found  only  exceptionally  in  human  lymph, 
but  frequently  in  calf  lymph.  These  included  a  bacterium  corre- 
sponding with  Proteus  vulgaris. 

Three  mice  were  inoculated  subcutaneously  with  a  drop  of  the 
liquefied  gelatine,  but  the  result  was  negative.  The  injection  of  a 
considerable  quantity  proved  fatal  to  guinea-pigs  and  rabbits,  a 
result  which  was  probably  due  to  ptomaine  poisoning. 

There  were  also  several  bacilli  which  did  not  liquefy  gelatine  ; 
these  were  not  investigated. 

Staphylococcus  cereus  albus  was  found  very  frequently,  and 
Staphylococcus  pyogenes  aureus  occasionally.  Pure-cultivations  of 
these  micrococci  inoculated  on  the  skin  of  calves  produced  a  rapid 
local  irritation,  followed  by  vesiculation,  but  without  the  classical 
characters  of  the  vaccine  vesicle.  The  inoculated  part  was  com- 
pletely healed  in  three  to  five  days.  According  to  Pfeiffer  they 
explain  the  so-called  false  vaccine. 

Micrococcus  pyogenes  albus  was  almost  constantly  present. 
Numerous  other  micrococci  were  found,  but  not  constantly  present ; 
vaccine  lymph  being  a  splendid  medium  for  the  growth  of  micrococci. 
Pfeiffer  pointed  out  that  the  effects  of  Staphylococcus  pyogenes  aureus, 
albus,  and  citreus,  and  of  Streptococcus  pyogenes  on  rabbits  had  an 
important  bearing  upon  the  practice  of  vaccination,  and  he  recom- 


INOCULATED   COW-POX.  325 

mended  that  calf  lymph  should  be  tested  before  use  upon  children 
by  inoculation  of  the  ear  of  a  rabbit.  If  after  two  days  no  erysipelas 
occurs  in  the  inoculated  rabbit,  the  absence  of  streptococci  may  be 
considered  as  almost  proved.  Two  or  three  rabbits  should  be  inocu- 
lated at  the  same  time. 

The  author's  researches  into  the  bacteriology  of  vaccine  lymph 
extended  over  some,  years.  They  independently  confirmed  and 
extended  the  results  obtained  by  Pfeiffer.  Having  on  several 
occasions  examined  vaccine  lymph  and  vaccine  pus,  and  failed  to  find 
a  specific  bacterium,  the  author  proceeded  to  make  a  more  systematic 
examination  of  the  different  species  of  bacteria  in  samples  of  current 
vaccine  lymph.  Pure-cultivations  were  obtained  by  plate-cultivation, 
and  inoculation  of  the  surface  of  nutrient  agar,  obliquely  solidified 
in  test  tubes.  Various  current  stocks  of  lymph  were  used  in  the 
investigation.  Among  the  specimens  of  calf  lymph,  No.  1  yielded  a 
torula,  Bacillus  pyocyaneus  and  Bacillus  subtilis;  No.  2,  a  bacterium,  a 
variety  of  proteus,  Staphylococcus  pyogenes  aureus,  and  yellow  bacte- 
rium ;  No.  3,  a  bacterium,  micrococcus,  yellow  bacterium,  and  torula  ; 
No.  4,  yellow  micrococcus,  white  micrococcus,  white  torula,  yellow 
sarcina,  white  diplococcus,  Staphylococcus  cereus  albus,  and  a  mould 
fungus;  No.  5,  yellow  sarcina,  Staphylococcus  pyogenes  aureus,  yellow 
micrococcus,  white  bacillus,  Staphylococcus  pyogenes  albus,  large 
white  micrococcus,  yellow  bacterium,  and  a  white  micrococcus.  Among 
the  specimens  of  human  vaccine  lymph,  No.  1  contained  a  white 
micrococcus,  proteus,  and  Staphylococcus  pyogenes  aureus ;  No.  2,  a 
micrococcus,  a  tetracoccus,  a  white  liquefying  micrococcus,  and  a 
yellow  bacterium ;  No.  3,  white  micrococcus,  yellow  micrococcus, 
Staphylococcns  aureus  and  flavus,  a  bacterium,  a  white  micrococcus, 
a  bacillus  resembling  Bacillus  subtilis,  Staphylococcus  pyogenes  cereus 
and  a  brown  tetracoccus.  The  author  is  familiar  with  these  different 
species  of  bacteria,  and  not  one  of  them  is  peculiar  to  vaccine  lymph  ; 
there  was  no  bacterium  constantly  present  in  human  and  calf  vaccine, 
and  there  was  not  one  which  could  be  regarded  as  the  contagium. 
To  sum  up,  most  of  them  are  well  known  saprophytic  bacteria,  and 
some  were  identical  with  bacteria  commonly  found  in  suppuration. 
Vaccine  lymph  is  a  most  suitable  cultivating  medium  for  micro- 
organisms, and  bacteria  invariably  got  access  to  the  contents  of  the 
vaccine  vesicle.  There  is  no  evidence  to  be  obtained  by  the  present 
methods  of  research  as  to  the  bacterial  nature  of  the  contagium  of 
vaccine  lymph.  Copeman  obtained  similar  results,  and  thus  con- 
firmed the  author's  conclusions. 

Klein  and  Copeman  have  also  observed   minute  bacilli  in  calf- 


326  INFECTIVE   DISEASES. 

lymph  and  in  variolous  lymph.  Numerous  attempts  to  cultivate  them 
in  nutrient  media  and  in  the  living  animal  failed  entirely,  and  the 
identity  of  the  bacilli  could  not  be  determined.  Pfeiffer,  Guarnieri, 
Monti,  Buffer,  and  Plimmer  have  drawn  attention  to  structures  in 
lymph,  which  they  believe  to  be  of  the  nature  of  parasitic  protozoa. 
These  bodies  have  been  studied,  more  especially  in  the  tissues.  They 
are  four  times  the  size  of  ordinary  micrococci,  and  are  found  in  the 
clear  vacuole  in  the  protoplasm  of  epithelial  cells.  Whether  they 
are  really  parasites  or  altered  anatomical  elements  has  not  been 
determined.  No  other  conclusion  can  be  drawn  from  all  these 
observations,  except  that  the  nature  of  the  contagium  of  cow-pox 
is  unknown. 

ORIGIN  OF  COW-POX. 

Jenner's  original  theory  was  that  cow-pox  .  was  derived  from 
"  grease,"  but  subsequently  he  distinguished  between  cow-pox,  a 
disease  peculiar  to  the  cow,  and  "  grease,"  a  disease  transmitted  to 
the  cow  from  the  horse,  and  the  mistake  of  confounding  these  two 
diseases  was  attributed  to  farmers  and  farriers.  Thus  he  wrote  : — 

"  From  the  similarity  of  symptoms,  both  constitutional  and  local, 
between  the  cow-pox  and  the  disease  received  from  morbid  matter 
generated  by  a  horse,  the  common  people  in  this  neighbourhood 
when  infected  with  this  disease,  through  a  strange  perversion  of 
terms,  frequently  called  it  the  cow-pox." 

Jenner's  theory  of  the  origin  of  cow-pox  has  been  discouraged ;  so 
also  has  the  view  of  its  being  a  "  spontaneous  "  disease  in  the  cow, 
though  Ceely,  after  many  years  of  research  in  the  Yale  of  Aylesbury, 
could  never  discover  the  probability  of  any  other  origin.  Both 
opinions  have  given  way  to  the  theory  that  cow-pox  is  small-pox 
transmitted  to  the  cow — an  opinion  advocated  by  Baron,  and 
supported  by  an  erroneous  interpretation  of  Ceely's  and  Badcock's 
variolation  experiments.  Thus  the  cow-pox  and  grease  of  farmers 
and  farriers  no  longer  attracted  attention  in  this  country,  and  as 
natural  cow-small-pox  has  never  been  discovered,  cow-pox  has  been 
credited  with  being  extinct. 

For  a  full  discussion  of  this  subject  the  reader  is  referred  to  the 
work  by  the  author  on  the  History  and  Pathology  of  Vaccination, 
but  the  variolation  experiments  alluded  to  will  be  briefly  mentioned. 

In  1801  Gassner  inoculated  eleven  cows  with  small-pox  lymph, 
and  succeeded  in  one  in  producing  phenomena  indistinguishable  from 
the  results  of  ordinary  vaccination  with  cow-pox,  and  children  were 
inoculated  from  the  cow. 


ORIGIN   OF   COW-POX.  327 

In  1828  Dr.  McMichael  reported  that  several  physicians  in 
Egypt  had  obtained  similar  results,  and  children  were  successfully 
"  vaccinated." 

In  1836  Dr.  Martin,  in  America,  inoculated  the  cow's  udder  with 
variolous  lymph,  and  by  inoculating  children  produced  an  epidemic 
of  small-pox  with  fatal  cases.  In  1839  Reiter  of  Munich,  after  fifty 
unsuccessful  attempts,  succeeded  in  producing  a  vesicle,  and  a  child 
inoculated  from  the  vesicle  contracted  small-pox. 

In  1839  Dr.  Thiele,  after  a  number  of  unsuccessful  attempts  to 
inoculate  cows  with  variolous  virus,  succeeded  in  producing  a  vesicle 
with  the  physical  characters  of  the  vaccine  vesicle,  and  from  it  a 
stock  of  lymph  was  raised  from  which  over  three  thousand  persons 
were  inoculated.  Thiele's  method  was  to  inoculate  the  udder  with 
lymph,  and  to  select  for  the  purpose  young  cows  which  had  recently 
calved  and  had  delicate  skins.  In  England  Ceely  succeeded  by 
inoculating  the  vulva  of  a  heifer.  One  of  the  punctures  developed 
into  an  enormous  vesicle,  which  was  undoubtedly  variolous.  His 
assistant  punctured  his  hand  with  the  lancet  which  had  been  used 
to  open  the  vesicle,  and  febrile  symptoms  appeared,  followed  by  an 
eruption  on  the  face,  neck,  trunk,  and  limbs,  at  first  papular,  then 
vesicular,  and  finally  pustular.  The  lymph  was  used  in  children, 
and  "  vaccine "  vesicles  were  produced.  One  child  suffered  from 
vomiting  delirium,  and  extensive  roseola,  but  there  was  no  eruption 
in  any  other  case. 

In  1840  Badcock  of  Brighton  inoculated  a  cow  successfully,  and 
later  succeeded  in  variolating  thirty-seven  out  of  two  hundred  cows 
upon  which  he  experimented. 

In  1847  variolation  of  the  cow  was  successfully  performed  at 
Berlin,  but  the  virus  produced  variola,  and  one  of  the  children 
inoculated  died  of  confluent  small-pox. 

In  1864  Chauveau  inoculated  seventeen  animals  with  virulent 
small-pox  lymph.  Very  small  papules  resulted,  and  the  virus  from 
the  papules  produced  variola  in  a  child,  which  was  infectious  to  others. 
Klein  in  this  country  until  recently  was  uniformly  unsuccessful. 
Yoigt,  Fischer,  King,  Eternod,  Haccius,  Hime  and  Simpson,  have 
all  succeeded  in  inoculating  cows  and  producing  variola-vaccine. 

The  results  of  these  experiments  have  been  very  generally  misin- 
terpreted, and  claimed  by  some  as  conclusive  evidence  of  the  identity 
of  cow-pox  and  small-pox.  Instead  of  the  vesicle  being  regarded  as 
the  most  attenuated  form  of  variola,  the  experimenters  are  said  to 
have  succeeded  in  producing  cow-pox. 

It  is  quite  true  that  they  produced  phenomena  indistinguishable 


328  INFECTIVE    DISEASES. 

from  the  phenomena  of  an  ordinary  vaccination,  but  that  does  not 
mean  that  they  produced  the  disease  cow-pox.  The  vesicle  which 
followed  the  inoculation,  whether  papular  or  vesicular,  was  small-pox. 
Ceely,  Badcock,  Yoigt,  and  others,  succeeded  in  ingrafting  the  cow 
with  small-pox,  and  when  suitable  lymph  and  suitable  subjects  were 
employed,  the  virus  was  so  attenuated  that  a  benign  vesicle  resulted. 
Similar  results  were  obtained  by  Sutton  and  Dimsdale,  and  identical 
results  by  Adams,  Guillou,  and  Thiele,  by  inoculating  the  human 
subject  with  variolous  lymph  without  first  ingrafting  the  disease  on 
the  cow. 

Vaccination  with  variola-vaccine  is  simply  a  modification  of  the 
Suttonian  system  of  small-pox  inoculation,  only  in  the  first  remove 
the  cow  is  substituted  for  the  human  subject.     All  those  who  were 
inoculated  with  Ceely's,    Badcock's,    or    Simpson's    variola-vaccine, 
were  not  in  the  usual  meaning  of  the  word  vaccinated ;  they  were 
not  inoculated  with  cow-pox  but  they  were  variolated,  and  in  such 
an   extremely  attenuated  form  that  the  persons  so  variolated   do 
not  convey  the  infection.     By  judicious  selection  it  is  thus  possible 
to  obtain  a  strain  of  lymph  from  variola  which,  by  direct  inoculation 
of  the  human  subject  or  by  first  inoculating  a  cow,  is  deprived  of 
infectious  properties,  and  produces  on  the  arm  the  physical  characters 
of  an  ordinary  vaccine  vesicle.     This  has  been  regarded  as  a  proof 
of  the  identity  of  small-pox  and  cow-pox,  but  it  is  not  so.     Variola, 
and  cow-pox  are  not  the  only  diseases  caused  by  a  virus  which  can 
be  attenuated  until  only  a  vesicle  is  produced  with  the  characters  of 
an  ordinary  vaccine  vesicle.     The  results  which  have  been  obtained 
with  the  virus  of  cattle  plague  and  of  sheep- pox  and  horse- pox  have 
been  given  in  previous  chapters  ;  and  no  one  would  urge  on  this 
account  that  human  small-pox,  cattle  plague,  cow-pox,  sheep-pox, 
and  horse-pox  are  all  manifestations  of  the  same  disease.     Cow-pox 
has  never  been  converted  into  human  small-pox,  and,  in  their  clinical 
history  and  epidemiology,  natural   cow-pox   and  human  small-pox 
are  so  different,  that  the  comparative  pathologist  is  no  more  pre- 
pared to  admit  their  identity  than  he  is  prepared    to  admit  the 
identity  of  cow-pox  and  sheep-pox,  or  small-pox  and  cattle  plague. 

Protective  Inoculation. — Whether  vaccination  of  all  heifers 
on  a  farm  would  protect  them  from  cow-pox  when  they  came  into 
milk  is  not  known,  the  duration  of  the  immunity  in  calves  afforded 
by  vaccination  having  not  been  determined.  Calves  undoubtedly 
have  an  immunity  after  vaccination,  lasting  for  some  weeks. 

In  1896  Beclere,  Chambon,  and  Meriard  experimented  upon 
the  immunising  power  of  the  serum  of  vaccinated  calves.  They 


COW-POX   AND   SMALL-POX.  329 

concluded  from  experiments  on  animals  and  children  that  the  serum 
of  a  vaccinated  calf  from  ten  to  fifty  days  after  vaccination  will  give 
immunity  against  inoculated  cow-pox.  They  further  stated  that, 
whereas  the  immunity  given  by  vaccination  in  the  ordinary  way 
is  not  complete  until  the  eighth  day,  the  immunity  obtained  by 
injection  of  the  immunising  serum  is  immediate.  The  serum  has 
also  been  credited  with  therapeutic  properties  and  has,  it  is  said, 
proved  efficacious  in  cases  of  small-pox. 

Jenner  believed  that  cow-pox  did  not  protect  against  itself  but 
protected  against  small-pox,  and  for  a  century  this  has  been  a  subject 
of  much  controversy.  The  reader  is  referred  to  the  Reports  and 
conclusions  of  the  Royal  Vaccination  Commission. 

Stamping-out  System. — It  would  undoubtedly  be  an  advan- 
tage if  cow-pox  were  scheduled  under  the  Contagious  Diseases  Animals 
Act.  Cow-keepers  and  dairy-men,  being  anxious  that  their  trade 
should  not  be  interfered  with,  very  commonly  conceal  the  existence 
of  the  disease,  and  perhaps  nothing  is  known  about  it,  unless  a  milker 
infected  from  the  cows  seeks  for  medical  advice.  The  contamination 
of  the  milk  with  lymph,  pus,  crusts,  and  sometimes  blood,  renders 
it  unwholesome,  and  therefore  precautions  ought  to  be  taken  to 
prevent  its  occurrence.  If  the  infected  cows  in  a  herd  are  the  last 
to  be  milked,  and  the  milker  washes  his  hands  after  the  milking,  the 
disease  will  not  spread. 


CHAPTER    XXIII. 

DIPHTHERIA. 

DIPHTHERIA  is  a  specific  infectious  disease,  especially  of  children, 
characterised  most  commonly  by  inflammation,  and  infiltration  with 
lymph  cells  and  fibrine,  of  the  mucous  membrane  of  the  fauces, 
pharynx,  larynx  and  trachea,  followed  by  necrosis  of  the  mucous 
membrane  and  the  formation  of  a  greyish-white  false  membrane, 
the  diphtheritic  membrane.  In  some  cases  a  diphtheritic  membrane 
forms  in  the  stomach,  intestine,  the  urinary  organs  and  in  wounds. 
After  the  separation  of  the  membrane  an  ulcer  remains,  which  may 
gradually  heal.  In  the  superficial  part  of  the  diphtheritic  membrane 
there  are  masses  of  bacteria  including  cocci,  streptococci,  and  bacilli. 
The  diphtheria  bacilli  are  not  found  in  the  blood  or  in  the 
internal  organs.  There  is  no  doubt  of  the  fact  that  diphtheria 
is  a  disease  which  can  be  communicated  from  one  person  to 
another ;  but  the  question  of  its  origin  is  still  a  vexed  one.  There 
is  a  close  association  with  insanitary  conditions  and  decaying 
animal  and  vegetable  refuse,  and  dampness.  Localities  with  damp 
houses,  defective  drainage,  and  a  cold  exposure,  are  favourable 
to  the  development  of  diphtheria;  but  that  does  not  necessarily 
indicate  that  these  conditions  can  originate  it.  On  the  other 
hand,  assuming  the  disease  to  be  due  to  a  living  contagium, 
these  insanitary  conditions  would  afford  a  suitable  environment 
predisposing  to  the  development,  and  facilitating  the  spread,  of  the 
disease.  Scarlet  fever  and  measles  predispose  to  diphtheria;  and 
defective  sanitary  conditions,  causing  sore  throat,  may  indirectly 
act  as  a  predisposing  cause.  A  great  many  cases  have  been  quoted 
to  illustrate  the  possibility  of  the  conveyance  of  diphtheria  by  milk, 
and  the  theory  which  best  harmonises  with  all  these  observations  is 
the  existence  of  a  specific  bacillus,  which  may  be  readily  transferred 
from  the  throat  of  the  diseased  to  the  healthy  ;  which  finds  also  in 
milk  a  suitable  soil  for  its  growth,  and  by  its  agency  may  be  trans- 
mitted to  the  consumer.  Such  a  bacillus  was  discovered  by  Loffler, 

330 


DIPHTHERIA. 


331 


and  may  be  easily 
obtained  from  the 
throat  of  diphtheritic 
patients  in  the  fol- 
lowing manner : — 

Culture  Outfit. — 
Steel  rods  like  or- 
dinary knitting 
needles,  about  six 
inches  in  length,  are 
beaten  out  or  rough- 
ened at  one  end,  and 
a  pledget  of  wool  is 
twisted  round  so  as 
to  form  a  swab. 
These  swabs  are 
placed  in  clean  test- 
tubes,  which  are  then 
plugged  with  cotton- 
wool. The  test-tubes 
and  swabs  are  steri- 
lised by  heating  in 
the  hot  air  steriliser 
f  or  an  hour  at  1 50°  C. 
The  so-called  culture 
o  u  t  fi  t  consists  of 
a  small  box  con- 
taining a  test-tube  of 
blood  serum  and  a 
swab.  They  can  be 
always  kept  ready  for 
use,  and  after  use 
should  be  conveyed 
by  hand  for  further 
examination.  The 
danger  of  trans- 
m  i  1 1  i  n  g  virulent 
diphtheritic  material 
by  post  is  obvious. 
When  the  examina- 
tion of  the  tube  has 
been  completed,  the 


f^'^^^^^^f^'^^^^f^ 

^S^M£SjtKI":r 


FIG.  126. — FREE  SURFACE  OF  DIPHTHERITIC  LARYNX 
x  350  (HAMILTON). — A,  Deposit  of  diphtheria  bacillus 
on  surface  of  false  membrane ;  B,  false  membrane ; 
(7,  mucosa ;  /,  lymph-cells  and  false  membrane 
surrounded  by  meshes  of  fibrine  ;  e,  surface  of  mucosa 
deprived  of  its  epithelium  ;  l,v,  lymph-cells  containing 
shed  epithelium. 


332  INFECTIVE    DISEASES. 

culture  outfit  and  its  contents  should  be  disinfected  or  destroyed. 
To  inoculate  the  tubes  the  patient,  if  it  is  possible,  should  be  turned 
to  the  light,  the  mouth  well  opened,  the  tongue  depressed,  and  the 
swab,  without  touching  the  teeth  or  the  tongue,  should  be  passed 
straight  to  the  tonsils  or  pharynx,  and  especially  to  the  membranous 
exudate.  The  swab  is  carefully  and  quickly  withdrawn,  and  at 
once  very  gently  rubbed  over  the  surface  of  the  blood  serum.  The 
culture  outfit  is  then  sent  to  the  laboratory  with  full  particulars, 
and  the  tubes  are  placed  in  the  incubator  at  37°  0.,  and  can  be 
examined  after  twelve  hours.  If  the  throat  has  been  disinfected 


IS 


S'SSl?  .-.!» 


V"W.-V*y     \  ^ 

^      "Wi  ^ 

x>\    ->•  *>-      " 


\t  »-\      • "   flr-f 

M'^t 

^^^•fc       »  -  » 


1  ^  A 

r  *- 

Wj,l     >f 

*H" 

ft  , 

^         ^ 
*  *&& 

*:-& 

f 

FIG.  127. — BACILLUS  OF  DIPHTHERIA  ;  FROM  A  CULTIVATION  ox  BLOOD 
SERUM,  x  1000  (FRANKEL  and  PFEIFFER). 

before  examination,  this  must  be  taken  into  account,  as  the  failure 
to  find  bacilli  would  not  then  necessarily  indicate  a  wrong  diagnosis. 
In  all  undoubted  cases  of  diphtheria,  growths  will  be  obtained  either 
in  the  form  of  a  pure-culture  of  the  bacillus,  or  far  more  commonly 
there  will  alsolbe  colonies  of  various  bacteria,  especially  Streptococcus 
pyogenes. 

Bacillus  of  Diphtheria — Rods,  straight  or  slightly  curved. 
•3  to  '8  /*  in  breadth,  and  1*5  to  6'5  ft  in  length.  They  occur 
singly,  in  pairs^  sometimes  in  chains,  and  sometimes  as  short 
leptothrix  forms.  In  some  cultures  very  irregular  forms  are 
observed,  the  bacilli  being  swollen  at  one  or  both  ends  or  thicker 
in  the  middle  portion,  or  the  bacillus  may  contain  oval  or  spherical 


DESCRIPTION    OF    PLATE    VIII. 
Bacillus  diphtherias  and  Bacillus  typhosus. 

FIG.  1. — Cover-glass  preparation  from  a  pure-cultivation  of   Bacillus   diph- 

therife  on  blood  serum  ;  obtained  from  the  throat  in  a  typical  case  of 

diphtheria.     Stained  with  gentian-violet,      x  1200. 
FIG.  2. — Cover-glass  preparation  from  a  pure-cultivation  of  Bacillus  typhosus 

on  nutrient-agar ;  from  the  spleen  in  a  case  of  typhoid  fever     Stained 

with  gentian-violet,      x  1200. 


Plate  MIL 


/•' » •  »ji  **      *•"••  *•% 
V  *    •',•»*  •*,         •••» 


,    t>*ol 

YcA-^&fe 


V» 


Fig  1.  BACILLUS    DIPHTHERIA 


. BACILLUS     TYPHOSUS 


DIPHTHERIA. 


333 


elements.  They  differ  greatly  in  size  and  shape,  often  in  the  same 
cultures,  and  still  more  in  cultures  obtained  from  different  sources. 
Spore  formation  is  unknown.  In  unstained  preparations  there  are 
highly  refractive  elements  which  correspond  with  the  deeply  stained 
parts  of  the  bacillus.  They  stain  readily  with  the  ordinary  aniline 
dyes.  At  certain  stages  of  their  growth  they  stain  irregularly,  the 
protoplasm  of  the  rod  being  broken  up  into  irregular  segments. 
The  bacillus  is  non-motile,  and  does  not  liquefy  gelatine  ;  it  grows 
at  20°  C.,  but  much  more  readily  at  higher  temperatures.  Colonies 
in  gelatine  plate -cultivations  are 
yellowish-brown,  and  opaque, 
granular,  and  circular,  but  with 
more  or  less  irregular  margin. 

In  plate -cultivations  on  agar 
and  on  glycerine  agar  the  same 
description  applies. 

On  the  surface  of  gelatine 
the  appearances  depend  greatly 
on  the  method  of  inoculation. 
The  growth  may  occur  in  the 
form  of  a  whitish  film,  but  if  a 
sub-culture  has  been  prepared 
from  broth  the  growth  is  often 
composed  of  a  number  of  iso- 
lated white  colonies  (Fig.  128,  a). 

On  blood  serum,  after  twelve 
hours  the  colonies  appear  in  the 
form  of  little  elevated  greyish - 
white  or  pearl-grey  dots,  which 
coalesce,  forming  a  film  if  the 
serum  is  moist.  On  the  surface 
of  1  per  cent,  alkaline  glycerine 
agar,  the  appearances  are  found 
to  vary,  and  this  medium  is  not  so  suitable  for  the  cultivation  of  the 
bacillus.  In  slightly  alkaline  broth,  with  or  without  the  addition  of 
1  per  cent,  grape-sugar,  the  culture  is  cloudy,  or  a  fine  granular 
deposit  occurs  along  the  sides  and  bottom  of  the  tube,  while  the 
broth  remains  clear. 

On  potato  the  growth  is  almost  invisible,  in  the  form  of  a  dry, 
thin  glaze.  Irregular  forms  are  very  numerous  on  microscopical 
examination,  whilst  the  rods  are  thicker  than  usual  (Welch  and 
Abbott).  In  milk  the  organisms  grow  readily. 


FlG.  128. — PUKK-CULTURES  OF  BACILLUS 

DIPHTHERIA  ON  GELATINE  :  a,  isolated 
colonies  ;  b,  filmy  growth. 


3.34  INFECTIVE    DISEASES. 

Dried  diphtheritic  membrane  and  cultures  dried  on  silk  threads 
retain  their  vitality  for  several  months. 

A  broth-culture  in  forty-eight  hours  may  be  used  for  inoculating 
guinea-pigs.  A  few  drops  will  cause  death  in  from  three  to  five 
days ;  there  is  hypersemia  and  oedema  at  the  seat  of  inoculation,  the 
lymphatic  glands  are  enlarged,  there  is  fluid  in  the  peritoneal, 
pleural  and  pericardial  cavities,  and  the  lungs  are  congested.  The 
bacillus  is  found  at  the  seat  of  inoculation,  but  not,  as  a  rule,  in  the 
blood  or  internal  organs.  Inoculation  of  rabbits  produces  extensive 
local  oedema,  enlargement  of  the  lymphatic  glands,  and  death  in 
from  four  days  to  three  weeks.  Roux  and  Yersin  pointed  out  that  in 
less  acute  cases  there  was  paralysis  of  the  hind  limbs.  Mice  and 
rats  have  an  immunity.  Cultures  lose  their  virulence  with  age,  but 
the  filtrate  from  old  cultures  contains  more  toxic  substance  than 
that  from  fresh  cultures.  The  toxin  has  been  described  in  a 
previous  chapter  (p.  46). 

Old  cultures  sterilised  by  heating  for  an  hour  to  60°  C.  or 
70°  C.  will  render  guinea-pigs  immune  in  two  weeks.  The  toxic 
substance  is  believed  to  be  destroyed  by  this  process,  while  according 
to  Frankel  the  immunity-giving  substance  which  is  also  present  in 
the  culture  is  not  affected. 

According  to  Behring's  researches,  the  blood  of  immune  animals 
contains  diphtheria  antitoxin,  consequently  the  blood  of  an  im- 
mune animal  is  capable  of  neutralising  the  toxic  properties  in  a 
filtered  culture,  not  only  in  the  living  animal  but  when  added  to  the 
culture  in  a  test-tube.  These  researches  led  to  the  employment  of 
the  serum  of  an  immune  animal  as  a  therapeutic  agent  in  the  treat- 
ment of  diphtheria  in  man  (p.  58). 

Bacteriological  Diagnosis. — The  diphtheritic  bacilli  are  not  only 
found  in  the  throat  while  the  lesions  exist,  but  they  are  found  after 
all  sign  of  the  disease  has  disappeared.  In  some  cases  they 
persist  for  a  few  days,  in  others  for  three  or  four  weeks,  and 
in  rarer  cases  they  have  been  found  several  months  afterwards. 
They  have  also  been  found  in  the  throats  of  persons  in  health, 
especially  of  those  who  have  been  in  contact  with  cases  of  diph- 
theria, such  as  healthy  children  in  infected  families  and  healthy 
nurses. 

The  bacilli  which  persist  in  the  throat  after  recovery  may  be 
virulent  up  to  the  time  of  their  disappearance,  or  they  may  gradu- 
ally become  attenuated,  and  entirely  lose  their  pathogenic  properties. 
The  value  of  a  microscopical  examination  as  an  aid  in  the  diagnosis 
of  diphtheria  has  been  considerably  exaggerated,  and  unless  the 


DIPHTHERIA.  335 

bacillus  when  isolated  is  tested  by  inoculation  the  test  may  prove 
to  be  entirely  fallacious. 

Loffler  and  Yon  Hoffman  both  found  bacilli  in  healthy  throats, 
and  thus  created  doubt  as  to  the  importance  of  the  Loffler  bacillus. 
Hoffman  found  this  bacillus  in  the  throats  of  twenty-six  out  of 
forty-five  individuals,  some  of  them  suffering  from  scarlet  fever, 
measles  or  some  other  non-diphtheritic  affections,  while  the  rest  were 
healthy.  The  bacilli  from  these  sources  showed  slight  differences 
in  morphological  and  cultural  characteristics,  and  Hoffman  was 
unable  to  decide  whether  these  bacilli  were  diphtheria  bacilli,  which 
had  become  harmless,  or  whether  they  were  accidental  epiphytes, 
belonging  to  a  closely  allied  but  different  species. 

Roux  and  Yersjn  confirmed  these  observations.  In  a  hospital 
for  children  in  Paris  without  any  question  of  the  existence  of 
diphtheria  they  found  the  so-called  pseudo- diphtheria  bacilli  in 
fifteen  cases  out  of  forty-five.  In  a  school,  in  a  seaside  place 
entirely  free  from  diphtheria,  the  same  bacilli  were  found  in 
twenty-six  out  of  fifty-nine  children.  They  were  also  found  in 
children  with  simple  sore  throats,  and  in  five  out  of  seven  cases 
in  measles.  Roux  and  Yersin  concluded  that  these  bacilli  were 
not  distinct  from  the  Loffler  bacillus.  There  were  slight  variations, 
but  there  was  no  constant  difference  except  in  their  pa.thogenic 
properties.  The  appearance  of  the  colonies,  the  growth  in  broth, 
and  the  peculiar  morphological  elements  showed  characters  common 
to  both,  and  there  was,  in  fact,  less  difference  than  there  is 
between  attenuated  anthrax  and  virulent  anthrax  in  form  and  in 
cultures;  but  inoculations  of  the  bacillus  did  not  cause  death, 
though  in  some  cases  in  guinea-pigs  there  was  marked  cedema  at 
the  seat  of  inoculation.  On  the  other  hand,  Loffler's  bacilli 
possess  different  degrees  of  virulence,  some  cultures  producing  only 
temporary  cedema,  while  others  cause  death  in  twenty-four  hours. 

Yirulent  diphtheria  bacilli  subjected  to  a  current  of  air  can 
in  two  weeks  be  deprived  of  their  virulence  partially,  and  in  four 
weeks  entirely.  Weakened  bacilli  can  be  raised  in  virulence  by  the 
simultaneous  injection  of  the  streptococcus  of  erysipelas,  but  bacilli 
deprived  of  their  virulence  and  bacilli  originally  non-virulent  cannot 
be  made  to  assume  virulent  properties.  Escherich  maintained 
that  they  could  be  distinguished  by  comparative  cultures ;  that  the 
pseudo-diphtheria  bacilli  made  broth  alkaline,  so  that  in  forty-eight 
hours  litmus  was  turned  red  by  Loffler's  bacilli  and  blue  by  the 
false  bacilli.  The  bacilli  themselves,  according  to  Hoffman,  are,  as 
a  rule,  shorter,  wider  and  more  uniform  in  size. 


336  INFECTIVE    DISEASES. 

Parke  and  Beebe,  with  a  view  to  clearing  up  this  question,  made 
cultures  from  three  hundred  and  thirty  healthy  throats.  They 
found  bacilli  of  three  varieties  :  bacilli  characteristic  in  growth 
producing  acid  reaction  in  broth,  but  having  no  virulence ;  bacilli 
not  characteristic  in  growth  producing  an  alkaline  reaction  in  broth, 
not  virulent ;  and  bacilli  producing  acid  reaction  in  broth  and 
virulent.  The  virulent  characteristic  diphtheria  bacilli  were  found 
in  eight  cases,  non- virulent  diphtheria  bacilli  in  twenty-four,  and 
non-virulent  false  diphtheria  bacilli  in  twenty-seven.  They  con- 
cluded that  the  eight  cases  must  have  been  in  contact  with  diphtheria, 
although  the  throats  were  healthy.  With  regard  to  the  bacillus 
in  the  twenty-four  cases  they  regarded  it  as  the  true  diphtheria 
bacillus  which  had  lost  its  virulence,  and  the  bacillus  found  in  the 
twenty-seven  cases  showing  differences  in  size  and  manner  of  staining 
and  the  reaction  produced  in  broth  was  properly  designated  pseudo- 
diphtheria  bacillus. 

DIPHTHERITIC  DISEASES  IN  ANIMALS. 

There  are  diptheritic  diseases  of  the  lower  animals  which  are  in 
some  respects  similar  to,  and,  some  observers  maintain,  identical 
with,  human  diphtheria. 

In  pigeons  there  is  a  disease  accompanied  with  the  formation  of 
false  membranes  associated  with  a  bacillus  described  by  Loffler. 

Bacterium  of  Diphtheria  of  Pigeons  (Bacillus  columbarum, 
Loffler). — Short  rods  with  rounded  ends,  mostly  in  irregular  masses. 
In  plate-cultivations  on  nutrient  gelatine  they  formed  whitish  patches 
on  the  surface,  and  compact,  ball-like  masses  when  embedded  in  the 
gelatine.  They  were  also  cultivated  on  blood  serum  and  potatoes. 
Subcutaneous  inoculation  of  a  pure- cultivation  produced  in  pigeons 
local  inflammation  and  necrosis  ;  inoculation  in  the  mucous  mem- 
brane of  the  mouth  gave  the  appearances  of  the  original  disease. 
Other  animals  were  only  locally  affected,  except  mice,  in  which 
characteristic  symptoms  and  death  resulted.  They  were  isolated 
from  the  diphtheritic  exudations  in  pigeons,  and  in  sections  were 
found  in  the  vessels  of  the  lungs  and  liver. 

A  similar  disease  is  known  to  attack  fowls,  and  there  are  also 
diseases  with  development  of  false  membranes  of  the  respiratory 
passages  in  horses,  cats  and  swine.  Outbreaks  of  these  diseases 
have  been  said  to  occur  in  times  of  prevalence  of  diphtheria  in 
man,  and  their  intercom municability  has  been  suggested. 

Dr.  Turner  supposes  that  diphtheria  in  man  originates  in  diseases 


DIPHTHERITIC    DISEASES   IN   ANIMALS.  337 

simulating  diphtheria  in  cats,  pigs,  and  horses ;  and  Klein,  who 
accepts  this  theory,  maintains  that  cats  suffer  from  genuine 
diphtheria,  and  that  after  death  the  lungs  are  found  full  of  grey, 
consolidated  lobular  patches,  and  the  kidneys  are  enlarged  and 
white. 

Human  diphtheritic  membrane  inoculated  subcutaneously  in  cats 
produces  a  painful  swelling  in  the  groin,  and  fever,  and  a  fatal 
termination  in  a  week.  The  subcutaneous  and  muscular  tissues  at 
the  seat  of  inoculation  are  hsemorrhagic  and  oedematous.  The 
internal  organs  are  congested,  and  in  the  kidneys  the  medulla  is 
congested,  while  the  cortex  is  fatty. 

A  recent  culture  produces  illness  in  twenty-four  hours,  a  painful 
tumour  forms  at  the  seat  of  inoculation,  and  death  ensues  in  from 
two  days  to  a  week.  Pneumonia  and  fatty  white  kidney  are  found 
after  death,  and  the  tissues  at  the  seat  of  inoculation  are  hsemor- 
rhagic,  and  in  parts  almost  gangrenous. 

Klein  found  that  diphtheritic  membrane  or  a  pure-culture  in- 
oculated into  the  cornea  after  removal  of  the  superficial  epithelium 
produced  ulceration,  and  in  two  cases  perforation  of  the  cornea r 
and  purulent  panophthalmitis.  Bacilli  were  again  recovered  from 
the  ulcer  similar  in  cultural  characters,  but  conspicuously  shorter 
and  thinner. 

An  epidemic  occurred  amongst  cats  at  the  Brown  Institution. 
Five  out  of  fourteen  died.  The  symptoms  were,  running  from  the 
eyes,  sometimes  a  muco-purulent  discharge,  sneezing,  coughing,  and 
pulmonary  trouble,  resulting  in  emaciation  and  death  in  from  one 
to  three  weeks.  Aften  death  lobular  pneumonia  and  large  white 
kidney  were  found ;  and  in"  one  case  a  diphtheritic  condition  of  the 
trachea,  preparations  of  which  showed  diphtheria  bacilli  in  crowds 
under  the  microscope. 

Klein  regarded  this  disease  as  an  epidemic  of  cat-diphtheria, 
and  believed  that  the  disease  was  possibly  induced  accidentally  by 
the  cats  drinking  milk,  which  was  infected  in  the  course  of  some 
other  experiments  with  diphtheria.  He  states  that  on  account 
of  the  very  definite  results  obtained  by  inoculating  diphtheritic, 
membrane  and  cultures  of  the  bacillus,  subcutaneously  and  on 
the  cornea,  and  of  the  condition  of  the  lung  and  kidney  in  cats 
naturally  or  experimentally  infected,  the  disease  must  be  con- 
sidered as  equivalent  to  human  diphtheria\A  and  the  cat  capable 
of  communicating  the  disease  to  other  cats,  and  also  to  human 
beings.  These  conclusions  have  not  yet  met  with  the  acceptance  cf 
veterinary  authorities.  The  results  of  the  experimental  inoculations 

22 


338  INFECTIVE   DISEASES. 

are  certainly  by  no  means  conclusive.  It  does  not  follow  from  these 
experiments  that  the  disease  diphtheria  naturally  occurs  in  the 
-cat  or  that  under  ordinary  circumstances  cats  may  contract  the 
disease  from  the  human  subject;  but  the  experiments  show  that, 
like  guinea-pigs  and  rabbits,  cats  are  susceptible  to  the  toxic  effects 
of  the  extremely  poisonous  principles  developed  during  the  growth 
of  Loffler's  bacillus. 


MILK  DIPHTHERIA. 

It  has  been  shown  that  milk  infected  with  diphtheria  has  been 
the  cause  of  epidemics  among  the  consumers ;  there  have  also  been 
epidemics  apparently  associated  with  the  milk  supply,  in  which  it 
has  not  been  possible  to  trace  the  source  from  which  the  milk  was 
infected.  A  difficulty  in  tracing  the  origin  in  no  way  excludes  the 
possibility  of  contamination  from  a  human  source.  In  the  light  of 
recent  researches  we  should  expect  that  it  would  be  easy  to  overlook 
the  source  of  the  virus,  if  it  be  true  that  diphtheria  may  exist  without 
.any  symptoms  indicating  its  presence,  and  be  unrecognised  until  the 
throat  has  been  examined  for  diphtheria  bacilli.  As  this  fact  was 
unknown  until  quite  recently,  the  absence  of  an  acknowledged 
•case  of  diphtheria  was  taken  as  evidence  that  no  diphtheria  existed, 
.and  consequently  that  the  milk  must  have  been  infected  by  a 
diseased  condition  of  the  cow.  Mr.  Power,  whose  views  upon  milk 
scarlatina  have  already  been  referred  to,  endeavoured  to  trace  the 
-origin  of  a  milk  epidemic  to  the  very  common  disease  of  "  garget," 
or  mammary  abscess.  This  idea  may  be  dismissed  without  further 
•consideration ;  but  the  theory  of  some  disease  existing  in  the  cow 
capable  of  producing  diphtheria  in  man  was  resumed  by  Dr. 
Cameron,  who  suggested  that  there  might  be  an  eruptive  disease  of 
the  teats  producing  diphtheria,  and  by  Mr.  Power,  who  supported  the 
theory  in  an  investigation  of  a  milk-diphtheria  outbreak  in  1886 
.at  Camber  ley.  Diphtheria  in  this  case  existed  in  the  neighbourhood, 
but  as  the  source  of  human  infection  could  not  be  traced,  attention 
was  drawn  to  two  cows  in  the  herd  which  had  recently  calved,  and 
•especially  to  one  with  chapped  teats.  Following  this  line  of  inquiry, 
Klein  investigated  the  behaviour  of  milch  cows  to  the  diphtheria 
bacillus.  Two  cows  were  injected  subcutaneously  under  the  skin  of 
the  shoulder  with  a  Pravaz' syringe  filled  with  a  sub-culture  in  broth. 
There  was  a  rise  of  temperature,  and  on  the  third  day  a  painful 
tumour,  which  enlarged  to  the  size  of  a  child's  head.  In  about  a 
fortnight  the  tumour  began  to  decrease,  and  ultimately  one  cow 


MILK   DIPHTHERIA.  339 

died  and  the  other  was  killed.  Such  results  might  have  been 
anticipated  as  the  result  of  injecting  a  large  quantity  of  the  toxic 
products  of  the  bacillus,  but  certain  other  phenomena  were  observed 
to  which  importance  was  attached.  On  the  fourth  day,  on  one  of 
the  cows  an  eruption  on  the  teat  was  first  noticed,  consisting  of 
small  vesicles  passing  into  pustules  and  crusted  ulcers.  Examina- 
tion of  the  contents  of  the  vesicle  revealed  the  bacillus.  With 
matter  from  the  vesicles  and  pustules  two  calves  were  inoculated, 
and  a  similar  vesiculation  produced  at  the  seat  of  inoculation. 
The  milk  of  the  cows  was  inoculated  on  nutrient  gelatine,  and 
produced  a  culture  of  Bacillus  diphtherise.  The  question  naturally 
arose  whether  this  eruption  had  any  connection  with  the  original 
experimental  inoculation.  No  other  cows  in  the  locality  from 
which  these  cows  were  obtained  had  a  similar  eruption,  and  it  was 
taken  for  granted  that  it  was  the  result  of  the  experimental  inocu- 
lation. By  accepting  the  possibility  of  this  eruption  being  identical 
with  the  chaps  on  the  teats  of  the  Camberley  cows,  the  theory  was 
gradually  built  up  that  cows  suffer  from  diphtheria,  which  manifests 
itself  in  the  form  of  an  eruptive  disease  of  the  teats,  and  that  the 
disease  is  conveyed  in  the  milk  to  the  consumers. 

In  the  original  experiment  the  bacilli  were  found  to  have 
multiplied  abundantly  in  the  tumour  at  the  seat  of  inoculation.  The 
eruption  might  have  been,  as  admitted  by  Klein,  a  symptom  of  the 
work  of  the  chemical  poison,  and  the  elimination  of  the  bacilli  by  the 
milk  is  also  possible ;  but  that  there  is  in  cows  a  vesicular  disease  of 
the  teats  which  is  the  origin  of  human  diphtheria  is  not  accepted  by 
veterinarians,  and  there  is  not  sufficient  evidence  to  justify  the  con- 
clusion that  the  infectivity  of  the  milk  in  epidemics  of  milk  diphtheria, 
has  been  proved  to  be  due  to  a  morbid  condition  of  the  cow. 


CHAPTER    XXIV. 

TYPHOID   FEVER. 

TYPHOID  FEVER  is  a  specific  febrile  disease  peculiar  to  man,  with 
characteristic  pathological  lesions  in  the  intestine,  mesenteric 
glands,  and  spleen.  The  Peyer's  glands  pass  through  three  stages. 
They  become  swollen  from  infiltration  of  round  cells  in  lymph 
follicles,  due,  it  is  supposed,  to  the  presence  of  the  typhoid  fever 
bacillus.  The  enlargement  of  the  lymph  follicles  is  followed  by 
coagulation  necrosis  until  the  entire  patch  becomes  necrosed,  and 
sloughs  away,  leaving  an  ulcer.  The  disintegration  of  the  patch 
may  extend  in  depth,  and  result  in  perforation  and  peritonitis,  or 
the  ulcer  may  heal,  and  a  pigmented  scar  take  the  place  of  the 
Peyer's  patch.  The  lymphatic  glands  are  found  more  or  less 
enlarged,  and  may  be  easily  felt  in  the  groin,  axilla,  and  neck.  In 
some  cases  there  is  a  tendency  to  haemorrhage,  followed  by  infarctions 
in  the  spleen  and  lungs,  which  may  develop  into  pysemic  abscesses 
In  the  mesenteric  glands  similar  changes  take  place,  but  without 
ulceration.  Pneumonia  may  occur  as  a  pulmonary  complication. 
The  bacteria  of  pneumonia  and  Streptococcus  pyogenes  may  be 
found  in  association  with  the  bacillus  of  typhoid  fever.  It  is 
now  generally  accepted  that  the  disease  is  conveyed  by  water  and 
food  which  have  become  contaminated  with  the  virus  contained 
in  typhoid  evacuations.  This  has  been  practically  proved  by  the 
number  of  cases  which  have  been  shown  to  have  been  intimately 
connected  with  contamination  of  drinking  water  from  wells  and 
other  sources,  by  sewers,  cesspools  and  faulty  drains,  the  sewage 
presumably  having  been  infected  with  typhoid  excreta  ;  but  whether 
sewage  independently  of  typhoid  contamination  can  originate 
typhoid  is  still  an  open  question.  Accepting  the  former  theory 
as  a  working  hypothesis,  we  must  assume  that  a  typhoid  fever 
bacillus  exists  in  the  intestinal  evacuations,  and  that  it  must  be  able 
to  retain  its  vitality  under  very  varying  conditions  until  it  gains 

310 


TYPHOID   FEVER. 


341 


access  by  the  mouth  to  a  fresh  host,  and  by  its  development  in  the 
intestine,  and  by  the  absorption  of  its  toxic  products,  produces  the 
phenomena  which  we  recognise  as  typhoid  fever. 


It 

'  -> , 


FIG.  129. — TYPHOID  FEVER.    ILEUM  OP  ADULT,  SHOWING  SLOUGHY  AND 
INFILTRATED  PATCHES  (HAMILTON). 

Typhoid  fever  is  also  disseminated  by  milk  ;  sewage- contaminated 
water  having  been  added  to  the  milk,  or  used  for  washing  the  milk 
cans  and  other  vessels. 


342 


INFECTIVE   DISEASES. 


Typhoid  fever  cannot  be  communicated  to  the  lower  animals. 
Numerous  experiments  have  been  made  by  feeding  and  by  injecting 
typhoid  stools,  but  with  absolutely  negative  results.  Murchison 
gave  typhoid  fever  discharges  to  pigs,  Klein  experimented  with 
rabbits,  monkeys,  and  other  animals.  Motschutkowsky  injected  the 
blood  from  cases  of  typhoid  into  monkeys,  rabbits,  and  other  animals, 
but  with  negative  results. 


FIG.  130.— TYPHOID  BACILLI  FROM  A  COLONY  ON  NUTRIENT  GELATINE,    x  1000 
(FRANKEL  AND  PFEIFFER). 

Various  micro-organisms  have  been  described  in  typhoid,  but  the 
one  to  which  most  importance  is  attached  is  a  bacillus  which  was 
first   discovered  by  Eberth,  but  cultivated   and  fully  described   by 
Gaff ky.    Gaffky  cultivated  it  from  typhoid 
evacuations,    from    typhoid    ulcers,    from 
the     mesenteric    glands,    and    from    the 
spleen.     It  is  found  in  scattered  colonies 
in  the  spleen,  and  is  rarely  if  ever  present 
in  the  blood. 

Bacillus  of  Typhoid  Fever.— Rods 
1  to  3  /x  in  length,  and  -5  to  -8  /A  in  breadth,  and  threads  (Plate  VIII., 
Fig.  2).  Spore-formation  has  not  been  observed,  but  the  protoplasm 
may  be  broken  up,  producing  appearances  which  may  be  mistaken 
for  spores.  They  are  actively  motile,  and  provided  some  with  a  single 
and  others  with  very  numerous  flagella,  which  are  from  three  to  five 
times  as  long  as  the  bacilli.  They  stain  well  with  aqueous  solutions 


FIG.  131.— TYPHOID  BACILLI, 
x  950  (BAUMGARTEN). 


TYPHOID   FEVER. 


343 


of  aniline  dyes,  and  grow  well  at  the  temperature  of  the  room.  In 
plate-cultivations  minute  colonies  are  visible  in  thirty-six  to  forty- 
eight  hours  ;  they  are  circular  or  oval,  with  an  irregular  margin  ;  they 
appear  granular  by  transmitted  light,  and  are  yellowish-brown  in 
colour.  Cultivated  in  the  depth  of  gelatine  a  well-defined  shiny  film 
forms  at  the  point  of  puncture,  and  a  greyish-white  filament,  com- 
posed of  closely  packed  colonies,  develops  in  the  track  of  the  needle 
(Fig.  134).  On  the  surface  of  gelatine  a  greyish-white  translucent 
film  forms,  with  sharply  defined  margin  (Plate  II.,  Fig.  2).  On  agar 
there  is  a  whitish  transparent  layer.  They  flourish  in  milk.  On 
potato  at  the  temperature  of  the  blood  there  is  no  culture  visible,  but 


FIG.  132.— FLAGELLA   OF    TYPHOID    BACILLI,    x    1000,    STAINED   BY  LOFFLER'S 
METHOD  (FRANKEL  AND  FFEIFFEB).  ' 

the  inoculated  area  appears  moist  and  shining,  and  cover-glass  pre- 
parations made  from  the  potato  will  demonstrate  that  there  is  really 
a  copious  growth  of  the  bacillus.  This  almost  invisible  growth  is  not 
peculiar  to  this  micro-organism. 

Whether  this  bacillus  is  really  peculiar  to  typhoid  is  much  dis- 
puted. Bacilli  very  closely  resembling  it,  if  not  actually  identical, 
have  been  found  under  other  conditions.  These  pseudo-typhoid  bacilli 
are  regarded  by  some  bacteriologists  as  varieties  resulting  from  the 
different  environment  afforded  by  a  saprophytic  existence.  Numer- 
ous experiments  have  been  made  on  animals  with  pure-cultures  of 
the  bacillus,  but  in  the  production  of  typhoid  fever  they  have 
been  no  more  successful  than  the  experiments  with  typhoid  stools. 


344 


INFECTIVE   DISEASES. 


Frankel  and  Simmonds  inoculated  a  number  of  rabbits  in  the  vein  of 
the  ear,  producing  death,  in  some  cases  .in  forty-eight  hours.  Seitz 
administered  broth-cultures  by  Koch's  method  of  introducing  them 

into  the  stomach  after 
the  administration  of 
opium  in  guinea-pigs, 
and  death  resulted  in 
several  instances. 
But  in  all  these  cases 
the  results  depended 
upon  the  poisonous 
products  found  in  the 


FIG.  133.— COLONIES  OF  TYPHOID  BACILLUS. 
Three  days  old.     x  100  (FRANKEL  AND  PFEIFFER). 


cultivations,  a  similar  result  following  the 
injection  of  sterilised  cultures.  An  account  of 
the  products  has  already  been  given  (p.  41). 

Cassedebat  isolated  three  species  of  bacilli 
from  water,  which  could  be  distinguished 
with  great  difficulty,  and  only  after  the  most 
careful  comparison.  The  bacillus  which  most 
closely  resembles  it  is  the  Bacillus  coli  com- 
munis;  in  fact,  Boux  regards  it  as  a  non- 
pathogenic  variety  of  the  typhoid  bacillus. 
Others  claim  to  be  able  to  distinguish  it  by 
careful  comparison  and  the  application  of 
tests.  Special  importance  is  attached  to 
potato  cultures,  the  typhoid  bacillus  forming 
an  invisible  film,  and  Bacillus  coli  communis 
a  well-marked  yellowish  growth.  Terni 
pointed  out  that  Bacillus  typhosus  retains  its 
motility  in  media  containing  hydrochloric 
acid,  while  Bacillus  coli  communis  and  other  bacilli  resembling  those 
of  typhoid  lost  their  motility.  Schild  maintained  that  Bacillus  typhosus 


FIG.  134. — PURE- CULTURE 
OF  TYPHOID  BACILLI 
INOCULATED  IN  THE 
DEPTH  OF  NUTRIENT 
GELATINE  (BAUMGAR- 
TEN). 


TYPHOID   FEVER.  345 

was  destroyed  by  exposure  to  the  vapour  of  formalin,  while  Bacillus 
coli  eommunis  and  similar  bacilli  isolated  from  water  gave  subcul- 
tures after  exposure  for  two  hours.  Typhoid  bacilli  do  not  give  the 
reaction  for  indol,  and  there  is  no  development  of  gas  in  cultures  in 
the  depth  of  nutrient  agar  containing  2  per  cent,  of  grape-sugar. 
According  to  Miiller,  sterilised  milk  is  coagulated  in  twenty-four 
hours,  at  37°  C.,  by  Bacillus  coli  eommunis,  but  not  by  the  Bacillus 
typhosus  until  several  weeks  have  elapsed ;  and,  further,  cultures  on 
acid  potato  give  different  results.  The  typhoid  bacillus  on  micro- 
scopical examination  shows  marked  polar  staining,  but  Bacillus 
coli  eommunis  only  shows  a  slight  tendency  of  the  protoplasm  to 
break  up. 


FIG.  135.— TYPHOID  BACILLI  IN  A  SECTION  OF  SPLEEN,    x   800  (FLUGGE). 

Kitasato  suggested  the  negative  indol  test,  and  recommended 
Salkowski's  method.  Broth-cultivations  are  treated  with  a  solution  of 
sodium  or  potassium  nitrite  :  1  cc.  of  the  nitrite  solution  (-02  gr.  to 
100  cc.  of  water)  is  added  to  10  cc.  of  a  broth-culture  after  twenty- 
four  hours  in  the  incubator,  and  on  adding  a  few  drops  of  strong 
sulphuric  acid,  the  typhoid  cultures  remain  colourless,  but  cultures 
of  bacilli  apparently  identical  give  the  red  colour.  On  the  other 
hand,  Losener  maintains  that  he  has  cultivated  from  earth,  water 
and  healthy  human  evacuations  bacilli  which  could  not  be  distin- 
guished from  typhoid  bacilli  by  any  of  these  tests. 

The    detection   of    the    typhoid    bacillus    in    water    has    been 


346 


INFECTIVE   DISEASES. 


described  in  another  chapter  (p.  147) ;  but  sufficient  has  been  said  to 
show  that  bacteriological  reports  in  which  it  is  stated  that  the  typhoid 
fever  bacillus  has  been  found  in  water  causing  typhoid  epidemics 
must  be  accepted  with  great  reserve  ;  and  further,  no  one  is  justified 
in  stating  that  the  typhoid  fever  bacillus  is  undoubtedly  the  cause 
of  typhoid  fever.  It  is  not  found  in  every  case  of  typhoid,  it  is 
not  found  in  the  blood,  but  it  is  found  in  those  tissues  which  are 


FIG.  136. — TYPHOID  BACILLI  IN  A  SECTION  OF  INTESTINE,  INVADING  THE  SUBMUCOUS- 
(T.J.)  AND  MUSCULAR  LAYERS  (M.),  x  950  (BAUMGARTEN). 

commonly    the   seat   of   secondary  invasion   of   epiphytic    bacteria, 
whose  normal  habitat  is  the  intestinal  canal. 

Lastly,  as  the  disease  does  not  exist  in  the  lower  animals,  the 
crucial  test  cannot  be  applied.  The  etiology  of  typhoid  fever  is 
still  enveloped  in  doubt,  and  the  nature  of  the  contagium  has  not 
yet  been  determined. 


CHAPTEK  XXV. 

SWINE   FEVER. 

PIG  TYPHOID,  or  swine  fever,  is  a  highly  contagious  disease  peculiar 
to  swine,  causing  death  in  from  ten  to  thirty  days,  associated  with 
a  nbrinous  pneumonia,  enlargement  of,  and  haemorrhage  into,  the 
lymphatic  glands,  and  characteristic  ulcers  of  the  mucous  membrane 
of  the  stomach  and  intestines.  The  lesions  may  assume  the  form  of 
extensive  croupous  or  diphtheritic  deposit,  which  may  fill  the  intes- 
tinal tube.  But  the  most  characteristic  appearance  results  when 
the  lower  part  of  the  ileum  and  commencement  of  the  colon  is 
dotted  all  over  with  elevations  of  the  mucous  membrane,  resembling 
leather  buttons  or  iiux  vomica  seeds,  and  sometimes  with  concentric 
rings,  so  that  they  have  been  compared  to  slices  of  calumba  root. 

Swine  fever  is  difficult  to  detect  in  the  early  stage,  and  sometimes 
symptoms  are  absent  altogether  in  animals  suffering  from  the 
disease  and  quite  capable  of  transmitting  it ;  or  nothing  may  be 
noted  except  cough,  and  possibly  enlargement  of  the  inguinal  glands. 
In  typical  cases  the  animals  are  noticed  not  to  feed,  to  exhibit 
dulness,  and  to  have  occasional  rigors.  Partial  paralysis  may 
follow,  producing  unsteady  gait  or  loss  of  power  over  the  hind  legs. 
Diarrhoea  sets  in,  and  the  evacuations  become  blood  stained.  There 
is  occasionally  a  diffused  or  patchy  reddish  or  purplish  rash  on 
the  skin.  After  death  the  appearances  most  commonly  found  are 
inflammation  of  the  peritoneum,  and  redness  and  enlargement  of  the 
mesenteric  glands  and  the  lymphatic  glands  in  the  lungs.  There 
is  generally  ulceration,  especially  of  the  colon  and  ileo-csecal  valve, 
or  a  diphtheritic  exudation,  sometimes  pale  yellow,  more  commonly 
greyish  or  black,  similar  to  the  centres  of  necrosis  within  the  ulcers. 
The  spleen  is  enlarged  and  liver  congested,  and  there  are  haemorrhages 
in  the  kidneys.  As  the  lungs  are  so  commonly  affected,  Klein 
proposed  the  name  pneumo- enteritis ;  but  the  pulmonary  lesions 
are  not  constant.  Indeed,  the  cases  in  which  the  intestines  and 

347 


348 


INFECTIVE   DISEASES. 


lungs  are  simultaneously  affected  are  not  numerous,  and  sometimes 
the   lungs   may  be  found   to  be   perfectly   healthy   in   cases   with 


PIG.  137.— ULCERATION  OF  THE  INTESTINE  IN  A  TYPICAL  CASE  OF  SWINE-FEVER. 

long-standing  lesion  of  the  intestine.  Old  pigs  may  linger  on  for 
weeks,  and  ultimately  recover,  and  in  the  meantime  act  as  centres 
for  the  dissemination  of  the  disease. 


DESCRIPTION  OF   PLATES   IX.   AND   X. 
Swine  Fever. 

PLATE  IX. — Part  of  intestine  from  a  typical   case  of  swine  fever,   showing 

scattered  ulcers  and  liberation  of  the  ileo-csecal  valve. 
PLATE  X.— From  the  same  case  of  swine  fever.     The  lungs  were  extensively 

inflamed  and  partly  consolidated,  and  the  lymphatic  glands  were  enlarged 

and  of  a  deep  red  or  reddish-purple  colour. 


Plate  IX. 


SWINE  FEVER 


K.  Croolulunk.ftei.1. 


Vincent  Brcoks.Day  &£o 


HateX 


SWINE     FEVER. 


Tfi.neen.1  BrocTca.Day  *Soa, 


SWINE   FEVER. 


349 


FIG.  138.— BACILLUS  OF  SWINE-FEVER 
No.  1.    (KLEIN.) 


Budd   first   pointed   out   that  this  disease   might   be   compared 

to   human   typhoid,    both   diseases   being   attended   by   a    peculiar 

ulceration  of  the  intestinal   follicles ;    but  the  diseases   are  not  to 

be  considered  in  any  sense  identical  or  interchangeable. 
Bacteria  in  Swine  Fever. 

—In    1877    Klein    published   a 

research  in  a  Report  to  the  Local 

Government     Board,    in     which 

he    claimed    to    have  discovered 

bacilli     characteristic      of      the 

disease.     They  were  described  as 

similar    to    Bacillus    subtilis,    or 

Bacillus    anthracis,    but    smaller 

in   size.     These  bacilli  developed 

into  long  leptothrix  filaments,  and  formed  spores.     It  was  further 

asserted  that  on  inoculation,  cultures  produced  lesions  indicative  of 

swine  fever  ;  the  bacilli  were  also  pathogenic  in  mice  and  rabbits. 

Later  this  bacillus  was  re- 
nounced in  favour  of  another. 
In  the  following  year  Det- 
mers  described  a  bacillus,  but 
subsequently  renounced  it  in 
favour  of  a  micrococcus. 

In  1882  Pasteur  maintained 
that  the  virus  of  swine  fever  in 
France  (rouget)  was  a  dumb-bell 
micrococcus,  which  produced  the 
same  effect  in  pigeons  as  the 
microbe  of  fowl -cholera.  Though 

rouget  or  swine  measles  is  probably  a  different  disease,  the  occurrence 

of  this  micro-organism  is  of  interest  in  this  connection. 

In   1883  Klein  again  investigated  swine  fever,  and   discovered 

Bacillus  No.   2,  and  maintained 

that  these  bacilli  were  found  in  ^  »*^=s 

£-j-frm  *  ^^ 

the  blood,  in  the  peritoneal  and  ^^  .?  « , 

bronchial  exudations ;  and  in  the  **      ^  «=»  % 

air  vesicles  of  the  lungs,  in  the 
form  of  leptothrix  filaments  ten 


, 


FIG.  139.— BACILLUS  No.  2.  FROM  A 
PREPARATION  OF  BRONCHIAL  Mucus 
OF  A  PIG.  (KLEIN.) 


FIG.  140.— BACILLUS  No.  2.    FROM  AN 
ARTIFICIAL  CULTURE.    (KLEIN.) 


or   twenty   times   the  length   of 

single   rods.      Cultivations  were 

made  on  solid  media.     The  organisms  in  these  cultures  were  minute 

rods  actively  motile,  occurring  singly  or  forming  chains,  two  or  three 


350 


INFECTIVE    DISEASES. 


times  as  long  as  Bacterium  termo ;  and  in  preparations  made  from 
diseased  organs  they  were  found  to  possess  a  very  narrow  trans- 
parent halo,  a  sort  of  hyaline  gelatinous  capsule.  Inoculation  of 
cultures  failed  to  produce  the  lesions  found  in  animals  naturally 
infected.  Two  pigs  were  inoculated,  one  with  a  sub-culture  from  the 
swollen  bronchial  gland  of  a  pig  that  had  died  of  pig-typhoid,  and 
a  second  with  a  culture  obtained  from  the  spleen  of  a  mouse  that 
had  been  inoculated  from  another  case  of  swine  fever.  After  two 
days  the  inguinal  glands  near  the  seat  of  inoculation  became  swollen, 
and  the  temperature  rose  slightly.  After  three  or  four  weeks  the 
animals  recovered. 

Mice  on  the  fifth  or  sixth  day  after  inoculation  showed  symptoms 

of  illness,  then  respiration  became 
superficial  and  slow.  Death 
occurred  on  the  sixth  or  seventh 
day. 

Rabbits  showed  a  rise  of 
temperature,  and  death  followed 
between  the  fifth  and  eighth  days, 
the  temperature  falling  before 
death.  At  the  post-mortem  ex- 
amination there  was  usually 
peritonitis,  with  copious  exuda- 
tion. The  kidney,  spleen,  and 
liver  were  enlarged  and  dark  in 
many  cases,  there  was  red  hepa- 
tisation  of  the  lobes  of  the  lungs, 
and  generally  pericarditis  and  haemorrhage  under  the  pericardium. 

In  1885  Salmon,  in  the  annual  report  of  the  United  States 
Bureau  of  Animal  Industry,  published  the  result  of  his  investigations 
into  American  hog  cholera,  which  is  identical  with  English  pig  typhoid. 
A  motile  figure-of-eight  bacterium  was  isolated,  each  part  being 
about  twice  as  long  as  broad.  The  bacterium  grew  on  nutrient 
gelatine  without  liquefying  it,  and  on  potato  produced  a  brownish 
growth  ;  broth  tubes  became  turbid  on  the  following  day.  Colonies 
in  plate- cultivations  were  oval  or  circular,  and  brownish  in  colour. 
Six  pigs  inoculated  subcutaneously  were  all  said  to  have  died  of  hog 
cholera,  and  the  bacterium  was  again  obtained  from  the  blood  of 
the  heart  and  spleen.  The  bacteria  proved  fatal  to  mice,  rabbits, 
guinea-pigs,  and  pigeons. 

In  1893  Welch  and  Clement  described  the  hog-cholera  bacillus 
as  variable  in  form,  and  they  further  stated  that  a  culture  obtained 


FIG.  141.— BLOOD  OP  FRESH  SPLEEN  OF 

A  MOUSE,    AFTER   INOCULATION    WITH 

(KLEIN.) 


BACILLUS  No.  2. 


SWINE   FEVER.  351 

from  Klein,  while  not  possessing  the  characters  originally  ascribed 
to  it,  could  not  in  its  form,  biological  characters,  or  pathogenic  pro- 
perties, be  distinguished  from  the  American  hog- cholera  bacillus. 

In  1887  pig  typhoid  was  investigated  at  Marseilles  by  Eietsch, 
Jobert,  and  Martinaud,  and  a  bacillus  found.  This  grew  rapidly 
on  all  the  nutrient  media.  In  gelatine  a  growth  was  obtained  in 
twenty-four  hours  at  18°  C. ;  on  blood  serum  and  agar  an  opaque 
growth  developed ;  and  on  potato  the  growth  was  yellowish.  It 
was  asserted  that  a  young  pig  was  killed  by  a  culture  in  twenty- 
two  days,  and  that  the  characteristic  ulcerations  were  observed  in 
the  intestines. 

In  1887  pig  typhoid  was  prevalent  in  Sweden,  and  Bang  and 
Selander  experimented  with  cultures  from  a  rabbit  that  died  after 
inoculation  with  a  fragment  of  spleen  from  a  diseased  pig.  The 
bacilli  were  motile,  varying  from  rods  to  cocci,  without  spore-formation 
and  pathogenic  in  mice,  guinea-pigs,  and  rabbits,  but  not  in  pigeons. 
Pigs  fed  on  broth -cultures  were  said  to  have  succumbed  to  genuine 
pig  typhoid.  In  the  blood  they  were  generally  found  in  the  form 
of  short  oval  bacteria,  but  in  the  blood  of  the  heart  longer  rods 
were  sometimes  found.  Metchnikoff  described  a  bacillus  isolated  by 
Chantemesse  from  an  outbreak  in  France,  as  highly  polymorphic. 

Smith  identified  the  hog-cholera  bacillus  with  the  bacillus  found 
by  Schiitz,  and  this  in  turn  has  baen  identified  with  the  bacillus 
of  haemorrhagic  septicaemia. 

From  these  researches  it  would  appear  to  be  probable  that  one  of 
the  bacteria  isolated  by  Klein,  and  those  found  by  Salmon,  Smith, 
Bang,  Welch  and  Schiitz  are  identical ;  and  further,  that  they  have 
been  identified  with  the  bacillus  of  hsemorrhagic  septicaemia.  We 
may  sum  up  the  characters  thus : — 

Bacillus  of  Klein,  Salmon,  Smith  and  Schiitz. — Very  small 
rods,  actively  motile  ;  spore-formation  not  observed  ;  colonies  circular 
and  brown  by  transmitted  light.  Inoculated  in  the  depth  of  gelatine 
faintly  yellowish- white  colonies  develop  along  the  track  of  the  needle  ; 
on  the  surface  an  opalescent  film ;  on  potato  they  produce  a  straw- 
coloured  layer,  changing  to  brown.  There  is  absence  of  indol  in 
cultures  containing  peptone;  the  bacilli  are  fatal  to  mice,  guinea- 
pigs,  rabbits,  and  pigeons.  Swine  die  after  intravenous  injection, 
but  not,  as  a  rule,  after  subcutaneous  injection. 

According  to  Caneva,  the  bacillus  obtained  from  the  Marseilles 
epidemic  would  appear  to  be  closely  allied,  if  not  identical,  with 
the  bacillus  of  ferret  disease  (Eberth  and  Schimrnelbusch),  and  the 
bacillus  of  Texas  fever  (Billings). 


352  INFECTIVE    DISEASES. 

Billings  appears  to  have  isolated  two  bacilli,  one  identical  with 
the  Marseilles  bacillus,  and  the  other  with  the  hog- cholera  bacillus. 

Bacillus  of  Rietsch  and  Jobert. — Rods  about  twice  as 
long  as  broad,  rather  shorter  than  the  bacillus  of  typhoid  fever, 
longer  and  thicker  than  the  American  bacillus.  They  exhibit 
end-staining.  They  possess  flagella,  and  are  actively  motile. 

They  grow  rapidly  in  nutrient  media.  They  are  only  feebly 
pathogenic.  They  are  also  said  to  be  distinguished  from  the  bacilli 
of  hog-cholera  by  producing  indol  in  solutions  containing  peptone, 
and  by  causing  an  acid  reaction  in  milk. 

Bacillus  of  M'Fadyean. — M'Fadyean  investigated  swine  fever 
in  1895,  and  found  bacilli  which  he  differentiated  from  hog-cholera 
bacilli.  The  method  employed  was  to  inoculate  the  surface  of 
nutrient  agar  and  of  potato,  with  fragments  torn  out  of  the  centre 
of  a  lymphatic  gland,  with  specially  constructed  forceps.  Inocula- 
tions were  also  made  from  the  spleen  pulp,  and  blood,  in  the  usual 
way.  They  are  from  1  to  2  /A  in  length,  and  '6  /*  in  breadth. 
They  never  grow  into  filaments,  they  do  not  form  spores,  and 
they  are  actively  motile.  They  are  readily  stained  by  the  watery 
solutions  of  the  aniline  dyes,  and  are  decolorised  by  Gram's 
method ;  with  methylene-blue  they  show  end-staining.  The  bacilli 
grow  on  gelatine  without  liquefaction,  forming  a  thin  white  line 
along  the  needle  track.  On  agar  a  thin,  transparent  pellicle  forms, 
which  is  not  easily  visible  at  first,  but  gradually  acquires  a  faint 
greyish  tint.  More  characteristic  appearances  result  in  plate- 
cultivations  of  gelatine-agar,  at  37°  C.  The  colonies  are  distinctly 
visible  in  eighteen  hours,  appearing  when  viewed  by  transmitted 
light  as  bluish-white,  circular  specks  ;  each  colony  has  a  dark  centre 
and  a  granular  margin.  In  broth  the  bacilli  produce  turbidity  after 
twenty-four  hours.  On  potato  there  is  no  visible  growth,  even  when 
the  surface  is  inoculated  with  an  abundance  of  material.  On  solid 
blood  serum  the  growth  is  scanty.  They  grow  in  milk  without 
producing  coagulation.  They  are  harmless  to  guinea-pigs  and  feebly 
pathogenic  to  rabbits. 

Several  experiments  were  carried  out  upon  swine.  In  the  first 
series  the  most  rigid  precautions  were  taken  to  prevent  accidental 
infection  with  swine  fever.  Four  young  pigs  were  inoculated,  upon 
a  farm  where  there  was  no  previous  history  of  the  disease.  These 
pigs  were  killed,  and  the  post-mortem  examinations  were  said  to 
show  indications  of  swine  fever,  principally  patches  of  diphtheritic 
material  in  the  colon,  and  healing  ulcers.  The  next  series  of  pigs 
was  inoculated  at  the  Royal  Veterinary  College.  Cultures  were 


SWINE   FEVER.  353 

administered  with  milk  to  two  pigs.  Five  days  afterwards  one  pig 
died ;  and  the  mesenteric  glands  were  congested,  and  the  mucous 
membrane  showed  spots  of  necrosis.  The  other  pig  was  killed,  and 
there  were  ulcers  in  the  colon.  In  a  third  experiment,  eight  pigs 
were  fed  with  milk  and  broth  cultures.  These  pigs  were  all  killed 
at  different  dates,  and  most  of  them  had  ulceration  of  the  colon  ; 
in  control  experiments  the  intestine  was  normal. 

M'Fadyean  compared  his  bacillus  with  a  culture  of  the  hog- 
cholera  bacillus,  and  found  that  the  American  organism  grows  at  a 
lower  temperature  in  gelatine,  and  colonies  appear  in  plates  much 
earlier.  They  produce  a  less  transparent  and  thicker  growth,  and 
much  greater  turbidity  in  broth  and  a  more  abundant  sediment. 
On  potato  they  form  an  abundant  growth  at  37°  C.,  at  first 
yellow,  later  brown,  with  considerable  resemblance  to  a  glanders 
culture. 

Colonies  upon  gelatine-agar  are  distinguished  by  their  opacity 
and  sharp  outline.  Agar,  potato,  and  broth  cultures  of  the  American 
organism  consist  of  short  ovoid  forms  like  the  bacilli  of  fowl  cholera, 
while  the  bacillus  isolated  by  M'Fadyean  has  a  closer  resemblance 
to  the  bacillus  of  glanders.  M'Fadyean  asserts  that  the  American 
organism  is  not  pathogenic  to  the  pig.  Pigs  after  feeding  on  broth 
cultures  remained  healthy,  and  showed  no  trace  of  swine  fever  when 
killed  from  one  to  three  weeks  afterwards.  On  the  other  hand, 
broth  cultures  of  his  bacillus  produced  the  characteristic  ulceration 
of  the  bowel.  M'Fadyean  claims,  therefore,  to  have  discovered  the 
true  pathogenic  organism  of  swine  fever.  He  does  not  appear  to 
have  compared  this  bacillus  with  that  obtained  from  the  epidemic 
of  swine  fever  at  Marseilles.  From  the  description  of  the  mor- 
phological and  other  details  there  seems  to  be  a  close  resemblance 
between  the  two. 

Not  less  than  three  and  possibly  four  species  of  bacilli  have  been 
cultivated  from  swine  fever,  two  at  different  times  by  Klein,  one 
by  Reitsch  Jobert  and  Martinaud,  and  one  by  M'Fadyean  ;  and 
cultures  of  all  these  bacilli  have  been  credited  with  producing  swine 
fever  in  experimental  animals,  and  each  one  has  been  pronounced 
to  be  the  contagium  of  the  disease.  We  must  conclude  either  that 
contaminated  cultures  were  inoculated  in  some  cases,  or,  what  is 
far  more  probable,  the  swine  fever  which  resulted  in  experimental 
animals  was  due  to  accidental  infection ;  and  until  a  bacillus  has 
been  cultivated  from  swine  fever  from  which  another  investigator 
can  prepare  sub-cultures,  and  with  those  sub-cultures  produce  the 
typical  u Iterations  of  swine  fever  in  pigs  on  a  farm,  or  on  premises 

23 


354  INFECTIVE    DISEASES. 

in  which  swine  fever  is  unknown,  we  are  justified  in  concluding  that 
the  contagium  has  not  yet  been  discovered. 

The  mistakes  which  are  likely  to  occur  when  the  same  investi- 
gator isolates  bacilli  from  cases  of  swine  fever,  and  subsequently 
inoculates  or  feeds  healthy  swine,  cannot  be  better  illustrated  than 
by  quoting  from  a  leaflet  issued  by  the  Board  of  Agriculture, 
pointing  out  the  great  precautions  necessary  to  prevent  accidental 
infection. 

"There  seems  reason  to  believe  that  the  disease  is  not  infre- 
quently introduced  by  means  of  persons  who  have  been  in  contact 
with  diseased  animals.  Pig  owners,  therefore,  are  advised  to  prevent 
strangers  from  at  any  time  approaching  their  pigs,  and  should  the 
admission  to  the  premises  of  spayers  or  castrators  be  necessary, 
those  persons  should  be  required,  before  approaching  the  animals,  to 
thoroughly  wash  their  hands  with  soap  and  water,  and  to  wash  and 
•disinfect  their  boots  with  a  solution  of  carbolic  acid  and  water,  or 
.some  other  suitable  disinfectant.  Such  persons  might  also  with 
advantage  be  required  to  wear,  while  operating,  a  waterproof  apron, 
which  should  be  washed  and  disinfected  before  the  wearer  is  per- 
mitted to  approach  the  animals  to  be  operated  on." 

Protective  Inoculation. — The  experiments  of  Salmon  and  of 
Schweinitz  have  been  referred  to  in  another  chapter  (pp.  41,  46). 
A  method  of  protective  inoculation  was  attempted  in  America,  but 
the  experiments  were  unsuccessful,  and  the  plan  was  abandoned. 

Stamping -out  System. — Notification  is  compulsory,  and  the 
order  in  force  is  the  Swine  Fever  Order  of  1896,  but  the  stamping- 
out  system  has  not  been  applied  in  a  thoroughly  satisfactory  manner, 
.and  the  disease  is  still  very  prevalent. 


CHAPTER    XXVI. 

SWINE    MEASLES. — DISTEMPER    IN    DOGS. — EPIDEMIC    DISEASE     OF 
FERRETS. — EPIDEMIC   DISEASE   OF   MICE. 

SWINE  MEASLES. 

SWINE  MEASLES,  or  swine  erysipelas,  is  described  as  an  acute,  in- 
fectious disease  of  swine  which  is  very  prevalent  in  France  and 
Germany,  but  is  included  in  this  country  in  the  term  "  swine  fever." 
According  to  some,  it  is  a  distinct  disease,  and  distinguished  from  pig- 
typhoid  by  absence  of  the  ulceration  of  the  intestines  which  is  so 
characteristic  of  that  disease  ;  while,  according  to  others,  ulceration 
of  the  intestine  and  ileo-ccecal  valve  may  be  found  post-mortem.  The 
onset  of  the  symptoms,  as  in  pig  typhoid,  is  very  rapid ;  the  animals 
cease  to  feed,  and  show  other  general  signs  of  illness ;  the  voice  is 
hoarse,  and  there  is  a  rapid  rise  of  temperature.  On  the  neck, 
chest,  and  abdomen,  red  patches  make  their  appearance,  which 
extend  and  coalesce,  and  change  to  a  dark  reddish  or  brownish  colour. 
These  symptoms  may  be  followed  by  convulsions,  and  sometimes  by 
paralysis  of  the  hind  legs;  and  death  occurs  in  from  one  to  four 
days.  It  is  especially  a  disease  of  young  pigs,  and  from  50  to  60 
per  cent,  of  infected  animals  die. 

On  post-mortem  examination  there  is  haemorrhage  and  oedema 
in  the  patches  of  the  skin,  the  lymphatic  glands  are  swollen  and 
dark  red,  the  peritoneum  is  ecchymosed,  the  intestinal  mucous 
membrane  is  congested  and  swollen,  and  the  solitary  follicles  and 
Peyer's  patches  are  prominent,  and  in  the  neighbourhood  of  the  ileo- 
ccecal  valve  there  are,  according  to  Fliigge,  ulcers  of  considerable  size. 
The  liver  and  spleen  are  congested  and  enlarged.  Pasteur  investi- 
gated swine  measles  or  rouget,  and  described  a  figure-of-eight  micro- 
coccus,  which  he  believed  to  be  the  contagium  of  this  disease.  This 
organism  appears  to  be  identical  with  the  bacterium  of  hsemorrhagic 
.septicaemia,  which  is  also  commonly  found  in  pig-typhoid. 

In  experimenting  with  the  virus  obtained  from  the  spleen 

365 


356  INFECTIVE    DISEASES. 

Pasteur  found  that,  by  successive  inoculation  of  rabbits,  the  virulence 
was  exalted  for  rabbits,  but  attenuated  for  swine,  and  the  virus 
which  had  thus  been  passed  through  the  rabbit  was  used  as  a 
vaccine  for  swine,  to  protect  them  against  virulent  erysipelas. 

Pasteur  found  that  by  passing  the  virus  through  pigeons  it  was 
made  more  virulent  for  swine. 

In  the  blood,  and  the  juice  of  the  internal  organs,  and  of  the 
lymph  glands,  Schiitz  found  a  minute  bacillus  identical  with  the 
bacillus  of  mouse  septicaemia. 

Bacillus  of  Swine  Erysipelas  (Schiitz).— Extremely  minute 
rods  '6  to  1'8  //.  in  length,  morphologically  and  in  cultural  charac- 
ters identical  with  the  bacillus  of  mouse  septicaemia.  Filaments 
and  involution  forms.  Spore-formation  present. 

House  mice  if  inoculated  with  a  pure  culture  die  in  two  to  four 
days.  Pigeons  are  also  very  susceptible.  Fowls  and  guinea-pigs 


FIG.    142.— BACILLI    OF    SWINE       FIG.  143.— BLOOD     OF    PIGEON     INOCULATED 
ERYSIPELAS  (BAUMGARTEN).  WITH  BACILLI  OF  SWINE  ERYSIPELAS,  x  600 

(SCHUTZ). 

are  immune.  Rabbits  after  inoculation  of  the  ear  suffer  from 
erysipelatous  inflammation,  identical  with  that  produced  by  inocu- 
lation of  the  bacillus  of  mouse  septicaemia.  The  bacilli  are  also 
pathogenic  in  swine  and  sheep. 

Protective  Inoculation. — With  Pasteur's  vaccine  immunity  is 
said  to  be  produced  which  lasts  about  a  year.  Schiitz  and  Schottelius 
found  the  minute  bacilli  in  Pasteur's  vaccine,  which  they  had  already 
found  in  cases  of  swine  erysipelas  in  Germany. 

The  results  of  vaccination  in  France  are  said  to  be  very  satis- 
factory, but  in  test  experiments  in  Germany  they  were  not  so 
favourable.  Out  of  119  vaccinated  swine.  5  per  cent,  died  as  the 
result  of  the  inoculation,  while  the  average  loss  in  the  ordinary  way 
is  2  per  cent. 

Metchnikoff  found  that  the  blood  of  immunised  rabbits  was 
antitoxic,  and  Lorenz  maintains  that  the  serum  of  swine  which 


SWINE   FEVER. 


357 


have  recovered  from  swine  erysipelas  is  also  antitoxic,  and  will 
produce  immunity  in  other  animals.  The  treatment  introduced 
by  Lorenz  is  to  inject  serum  in  the  proportion  of  1  cc.  to  every 
10  kilogrammes  of  the  weight  of  the  animal's  body.  Two  days 
afterwards  -5  to  1  cc.  of  virulent  culture  is  injected,  and  after 
twelve  days  the  dose  is  doubled.  Lorenz  inoculated  294  pigs ; 
12  were  suffering  from  swine  erysipelas,  and 
of  these  6  recovered  and  6  died. 

In  the  opinion  of  the  author  this  disease 
requires  re-investigation,  for  if  it  be  true  that 
rouget  or  schweinrothlauf  is  associated  with 
ulceration  of  the  intestines,  the  recognition 
of  it  as  a  disease  distinct  from  our  English 
swine  fever  apparently  rests  upon  the  pres- 
ence of  a  bacillus,  which  cannot  be  distin- 
guished from  the  bacillus  of  mouse  septicaemia. 
The  question  arises  whether  this  bacillus 
is  really  the  cause  of  a  distinct  disease,  swine 
erysipelas,  or,  on  the  other  hand,  whether 
the  bacillus  is  really  the  bacillus  of  mouse 
septicaemia  which  has  been  isolated  from 
certain  cases  of  swine  fever.  The  bacillus  of 
mouse  septicaemia  is  widely  distributed,  and 
it  may  only  be  an  accidental  concomitant  in 
rouget  or  schweinrothlauf.  The  presence  of 
the  bacterium  of  haemorrhagic  septicaemia  in 
both  rouget  and  pig  typhoid  would  not  prove 
identity,  as  this  micro-organism  is  un- 
doubtedly only  secondary  in  both  diseases. 
There  is  great  need,  therefore,  for  further 
careful  investigation.  Clinical  and  patho- 
logical observations  must  be  made  in  this 
country,  to  determine  whether  there  are 
really  two  diseases  included  under  the  name 

"  swine  fever."  If  this  prove  to  be  the  case,  we  must  ascertain  the 
clinical  and  pathological  differences  between  rouget  and  pig  typhoid. 
How  can  rouget  be  distinguished  from  cases  of  swine  fever  in  which 
there  is  a  patchy  rash,  paralysis  of  hind  legs,  but  no  ulceration  of  the 
intestine  ?  Further,  how  is  swine  erysipelas  with  ulceration  of  the 
intestine  and  ileo-ccecal  valve  to  be  distinguished  from  an  ordinary 
case  of  pig  typhoid  ? 


FIG.  144.— PURE-CULTURE 
IN  NUTRIENT  GELA- 
TINE OF  BACILLI 
IN  SWINE  ERYSIPELAS 
(BAUMGARTEN). 


358  INFECTIVE   DISEASES. 

DISTEMPER  IN  DOGS. 

Distemper  is  an  infectious  febrile  disease  of  dogs,  characterised  by 
bronchial  catarrh  and  discharge  from  the  eyes.  Bronchitis  and 
pneumonia  may  supervene,  or  there  may  be  intestinal  catarrh  ter- 
minating in  dysenteric  diarrhoea,  sometimes  complicated  by  jaundice. 
The  disease  may  affect  the  nervous  system,  and  produce  convulsive 
contractions  of  the  muscles  of  the  nose,  ears,  lips,  and  limbs. 
Occasionally  there  is  an  eruption,  especially  in  animals  which  are 
out  of  condition.  The  virus  exists  in  the  discharge  from  the  nostrils 
and  eyes,  and  is  given  off  from  the  lungs  and  the  skin. 

One  attack  of  the  disease  does  not  confer  entire  immunity ; 
and  some  dogs  are  completely  insusceptible. 

Bacteria  in  Distemper. — Millais  has  isolated  a  micro-organism 
resembling  the  pneumococcus  of  Friedlander,  which  he  believes  to  be 
the  cause  of  the  disease.  The  bacillus  occurs  with  other  bacteria  and 
micrococci  in  the  nasal  discharge. 

Protective  Inoculation. — Mixed  cultures  of  these  bacteria 
liquefy  the  gelatine,  and  the  liquid  has  been  used  as  a  vaccine. 
When  applied  to  the  nose,  it  is  said  to  produce  a  mild  attack  of 
distemper,  which  protects  as  much  as  an  attack  of  the  disease 
contracted  naturally.  These  results  require  confirmation. 

Inoculation  of  the  nasal  discharge  in  healthy  dogs  has  been 
practised,  so  that  they  may  have  the  disease  under  favourable  con- 
ditions ;  but  the  system  should  not  be  encouraged,  as  dogs  need  not 
necessarily  contract  distemper..  Vaccination  with  cow-pox  lymph 
has  been  advocated,  but  it  is  perfectly  useless. 

Stamping-out  System. — Dogs  suffering  from  distemper  must  be 
completely  isolated.  Any  straw  or  litter  which  has  been  in  contact 
with  a  diseased  dog  should  be  burnt.  Clothing,  collars,  chains,  and 
the  kennel  or  premises  inhabited,  must  be  thoroughly  disinfected. 
The  animal  after  recovery  should  be  washed  with  carbolic  soap. 

EPIDEMIC  DISEASE  OF  FERRETS. 

Eberth  and  Schimmelbusch  investigated  an  epidemic  disease  of 
ferrets  (frettchen-seiiche),  and  isolated  a  bacillus,  which  in  mor- 
phology and  cultivation  agrees  very  closely  with  the  bacillus  of 
hsemorrhagic  septicaemia. 

EPIDEMIC  DISEASE  OF  MICE. 

Lbffler  investigated  an  epidemic  disease  which  occurred  in  mice 
kept  in  confinement,  and  isolated  a  bacillus  resembling  Bacillus 
typhosus. 


EPIDEMIC   DISEASE   OF   MICE.  359 

Bacillus  Typhi  Murium. — Rods  varying  in  length ;  and  fila- 
ments ;  motile ;  flagellated.  The  colonies  are  circular,  brownish 
and  granular  on  the  surface  of  obliquely  solidified  gelatine.  The 
bacteria  inoculated  on  the  surface  produce  a  greyish-white  semi- 
transparent  growth,  and  on  agar  and  potato  the  appearance  of  the 
growth  is  very  similar.  They  can  be  cultivated  readily  in  milk  and 
in  broth.  White  and  field  mice  are  killed  in  from  one  to  two 
weeks,  when  given  bread  moistened  with  a  culture. 

Lofner  claims  to  have  used  this  method  with  success  in  Thessaly, 
where  there  was  a  plague  of  field  mice  causing  great  losses  to 
agriculturists. 


CHAPTEK    XXVII. 

ASIATIC  CHOLERA. — CHOLERA  NOSTRAS. — CHOLERAIC  DIARRHCEA 
FROM  MEAT-POISONING. — DYSENTERY. — CHOLERAIC  DIARRHCEA 
IN  FOWLS. 

ASIATIC  CHOLERA. 

THERE  are  several  diseases  in  man  associated  with  diarrhoea,  which 
have  certain  characters  in  common,  but  are  totally  distinct.  They 
include  Asiatic  cholera,  cholera  nostras,  dysentery,  and  choleraic 
diarrhoea.  Asiatic  cholera  is  an  endemic  disease  of  the  Delta  of 
the  Ganges,  a  locality  which  has  become  notorious  as  the  home 
of  cholera.  Cholera  is  a  filth  disease ;  and  the  accumulation  of 
filth  on  the  banks  of  the  Ganges,  with  contamination  of  the 
water,  and  the  climate,  afford  most  favourable  conditions  for  the 
development  of  the  cholera  virus. 

Four  great  cholera  epidemics  have  originated  in,  and  spread 
from,  India :  in  1817,  in  1826,  in  1846,  and  in  1865.  Cholera 
follows  the  routes  of  pilgrims  and  caravans,  and  now,  owing  to 
the  rapid  means  of  communication  by  steamers  and  railways,  it 
spreads  to  the  most  distant  parts  of  the  world,  covering  in  a  few 
weeks  or  days  distances  which  in  former  times  could  only  be  traversed 
in  several  months  or  even  years. 

In  1892  the  epidemic  passed  from  India,  through  Afghanistan, 
to  Russia  in  Asia,  and  quickly  spread  westwards  along  the  route 
of  the  trans-Caspian  railway ;  and  all  this  occurred  within  the  space 
of  a  few  weeks.  By  Russian  emigrants  it  was  carried  to  Hamburg 
and  Antwerp ;  and  the  virus,  finding  a  suitable  environment  in  the 
former  place,  produced  a  severe  epidemic  there.  Thus,  in  about  three 
months,  it  was  brought  into  close  proximity  with  England.  Mecca 
is  one  of  the  great  infective  centres  of  the  world,  for  there  all 
the  conditions  are  found  for  the  propagation  of  cholera,  including 
filth,  overcrowding,  and  the  water  of  the  famous  Holy  Well,  which 
is  used  for  ablutions  and  drinking  purposes.  The  return  of 

860 


ASIATIC   CHOLERA.  361 

the  pilgrims  to  Egypt,  and  the  proximity  of  England  to  Egypt, 
necessitate  the  greatest  possible  precautions  to  prevent  the  intro- 
duction of  the  disease  into  this  country. 

In  1884  a  German  Commission  was  sent  out  to  India,  and  Koch 
discovered  a  micro-organism  which  he  described  as  a  curved  or 
comma-shaped  bacillus,  and  pronounced  to  be  the  contagium  of 
this  disease. 


FIG.  145.— COVER-GLASS  PREPARATION  OF  A  DROP  OF  MEAT  INFUSION,  containing 
a  pure-cultivation  of  comma-bacilli,  with  (a)  spirilliform  threads,  x  600.     (Kocn.) 

Spirillum  choleras  Asiatics  (Comma-bacillus,  Koch). — Curved 
rods,  spirilla,  and  threads.  The  curved  rods  or  commas  are  about 
half  the  length  of  a  tubercle-bacillus.  They  occur  isolated,  or 
attached  to  each  other  forming  S~snaPecl  organisms  or  longer 
screw-forms,  the  latter  resembling  the  spirilla  of  relapsing  fever. 


FIG.  146.— ARTHROSPORES  ;  («)  Comma-bacillus  breaking  up  into  spheres;  (b,  c), 
formation  of  spheres  in  spiral  forms  ;  (d,  e\  groups  of  spheres ;  (/)  spirilla 
with  spheres  from  an  old  cultivation;  (ff)  germination  of  the  spheres. 

(HUEPPE.) 

Finally  they  may  develop  into  spirilliform  threads.  In  old  cultiva- 
tions threads  are  found  with  swellings  or  irregularities  (Fig.  148). 
The  commas  are  actively  motile,  and  possess  flagella  (Fig.  147). 
Their  movements)  and  development  into  spirilla  may  be  studied  in 
drop- cultivations.  Arthrospore  formation  has  been  described  by 
Hueppe  (Fig.  146).  In  plate -cultivations,  at  a  temperature  of  from 


362  INFECTIVE    DISEASES. 

16d  to  20°  C.,  the  colonies  develop  as  little  specks,  which  begin  to 
be  visible  after  about  twenty-four  hours.  Examined  with  a  low 
power,  and  a  small  diaphragm,  these  colonies  have  the  following 
characteristics.  They  appear  as  little  masses,  granular,  and 


FIG.  147. — FLAGELLA  OF  COMMA-BACILLI  ;  STAINED  BY"; LOPFLER'S  METHOD 
(FRANKEL  '  AND  PFEIFFER). 

yellowish-white  in  colour,  and  sometimes  very"-  faintlyi  tinged  « with 
red,  which  have  liquefied  the  gelatine,  and  sunk  down  to  the  bottom 
of  the  resulting  excavations. 

In  test-tubes  of  slightly  alkaline  nutrient  gelatine  (10  per  cent.), 


/ Of, 


FIG.  148.— INVOLUTION  FORMS,  x  700       FIG.  149.— COLONIES  OF  COMMA-BACILLI 
(VAN  ERMENGEM).  ON   NUTRIENT   GELATINE,    NATURAL 

SIZE  (KOCH). 

the  appearance  of  the  growth  is  very  striking.  In  typical  cultures 
it  begins  to  be  visible  in  about  twenty-four  hours.  Liquefaction 
sets  in  very  slowly,  commencing  at  the  top  of  the  needle  track 


ASIATIC   CHOLERA. 


363 


around  an  enclosed  bubble  of  air,  and  forming  a  funnel  continuous 
with  the  lower  part  of  the  growth  ;  the  latter  preserves  for  several 
days  its  resemblance  to  a  white 
thread  (Plate  II.,   Fig.   1).      In 
about  eight  days,  however,  lique- 
faction   takes    place    along   the 
whole  of  the  needle  track. 

On  the  surface  of  agar-agar 
the  cultivation  develops  as  a 
white,  semi-transparent  layer, 
with  well-defined  margin.  The 

appearance  on  blood  serum  is  very  similar ;  liquefaction  very  slowly 
takes  place.  In  broth  they  form  a  wrinkled  film  on  the  surface, 
there  is  a  rapid  and  abundant  growth  at  the  temperature  of  the 


FIG.  150. 


COLONIES  OF  KOCH'S  COMMA- 
BACILLI,   x  60. 


FIG.  151.— COVER-GLASS  PREPARATION 
FROM  THE  CONTENTS  OF  A  CHOLERA 
INTESTINE,  x  600.  (a)  Remains  of  the 
epithelial  cells;  (b)  Comma-bacillus; 
(c)  Group  of  comma-bacilli  (Koch). 


FIG.  152.— COVER-GLASS  PREPARATION 
OF  CHOLERA  DEJECTA  ON  DAMP  LINEN 
(two  days  old),  x  600.  Great  prolife- 
ration of  the  bacilli  with  spirilla  (a) 
(Koch). 


blood,  and  the  same  applies  to  sterilised  milk ;  and  they  will  even 
multiply  in  sterilised  water.  In  potato- cultivations  the  microbe 
will  only  grow  at  the  temperature  of  the  blood  (37°  C.),  forming  a 
slightly  brown,  transparent  layer.  Inoculation  of  a  cultivation  of 
the  bacillus  in  the  duodenum  of  guinea-pigs,  with  and  without 


364  INFECTIVE   DISEASES. 

ligation  of  the  bile-duct,  has  given  positive  results.  More  recently 
these  results  have  been  confirmed  by  the  following  method  :  Five 
cc.  of  a  5  per  cent,  solution  of  potash  were  injected  into  the 
stomach  of  a  guinea-pig,  and  twenty  minutes  after,  10  cc.  of  a 
cultivation  of  comma-bacilli,  diffused  in  broth,  were  similarly  intro- 
duced. Simultaneously  with  the  latter,  an  injection  of  tincture  of 
opium  was  made  into  the  abdominal  cavity,  in  the  proportion  of 
1  cc.  for  every  200  grammes  weight  of  the  animal.  Those  who 
have  had  success  with  inoculation  experiments  maintain  that  choleraic 
symptoms  were  produced  without  any  trace  of  peritonitis  or  putrid 
infection,  and  that  the  comma -bacilli  of  Koch  were  again  found 
in  the  intestinal  contents,  and  fresh  cultivations  established. 


FIG.  153.— SECTION  OF  THE  Mucous  MEMBRANE  OF  A  CHOLERA  INTESTINE,  x  600. 
A  tubular  gland  (a)  is  divided  transversely  ;  in  its  interior  (6)  and  between 
the  epithelium  and  the  basement  membrane  (c)  are  numerous  comma-bacilli 
(Koch). 

On  the  other  hand,  these  results  have  been  disputed,  the  fatal 
effects  of  the  inoculation  attributed  to  septicsemic  poisoning,  and 
the  proliferation  of  the  bacilli  considered  to  be  dependent  upon  an 
abnormal  condition  of  the  intestines,  induced  by  the  injection  of 
tincture  of  opium.  It  has,  however,  been  shown  that  these  organisms, 
like  several  others  which  have  been  isolated  from  intestinal  dis- 
charges, produce  definite  poisonous  substances.  The  comma-bacilli 
were  found  in  the  superficial  necrosed  layer  of  the  intestine,  in 
the  mucous  flakes  and  liquid  contents  of  the  intestinal  canal  of 
cases  of  Asiatic  cholera.  It  is  stated  that  they  were,  also  detected 


ASIATIC   CHOLERA.  365 

in  a  tank  which  contained  the  water  supply  of  a  neighbourhood 
where  cholera  cases  occurred;  but  comma-shaped  organisms  are 
frequently  present  in  sewage-contaminated  water.  Koch's  comma- 
bacilli  are  aerobic,  and  their  development  is  arrested  by  deprivation 
of  oxygen.  They  are  destroyed  by  drying  on.  a  cover  glass,  but 
retain  their  vitality  longer  when  dried  on  silk  threads.  Cultures 
are  sterilised  by  exposure  for  fifteen  minutes  to  55°  C.,  and  by 
various  antiseptic  substances. 


FIG.  154.— PURE-CULTIVATIONS  IN  NUTRIENT  GELATINE,  a,  KOCH'S  CHOLERA 
BACILLUS,  twenty-four  hours  old.  I,  TINKLER'S  BACILLUS,  twenty-four 
hours  old. 

METHODS  OF  STAINING  THE  COMMA-BACILLI  OF  KOCH. 

In  cover-glass  preparations  they  may  be  well  stained  in  the  ordinary 
way,  with  an  aqueous  solution  of  methyl-violet  or  fuchsine,  or  by  the 
rapid  method,  without  passing  through  the  flame  (p.  85,  Babes'  method). 

Nicati  and  Reitsch's  method. 

A  small  quantity  of  the  stools,  or  of  the  scraping  of  the  intestinal 
mucous  membrane,  is  spread  out  on  a  glass  slide  and  dried,  then  steeped 
during  some  seconds  in  sublimate  solution,  or  in  osmic  acid  (1  to  100). 
It  is  then  stained  by  immersion  in  fuchsine-aniline  solution  (1  or  2 
grammes  of  Bale  fuchsine  dissolved  in  a  saturated  aqueous  solution  of 
aniline),  washed,  dried,  and  mounted  in  Canada  balsam. 


366  INFECTIVE   DISEASES. 

In  sections  of  the  intestine  their  presence  may  be  demonstrated  by  :  — 

(a)  Koch's  method. 

Sections  of  the  intestine,  which  must  be  well  hardened  in  absolute 
alcohol,  are  left  for  twenty-four  hours  in  a  strong,  watery  solution  of 
methylene-blue,  or  for  a  shorter  time  if  the  solution  is  warmed  ;  then 
treated  in  the  usual  way. 

(6)  Babes'  method. 

Sections,  preferably  from  a  recent  case  of  cholera,  and  made  as  soon  as 
possible  after  death,  are  left  for  twenty-four  hours  in  an  aqueous  solution 
of  f uchsine,  then  washed  in  distilled  water,  faintly  acidulated  with  acetic 
acid,  or  in  sublimate  solution  1  in  1000,  passed  rapidly  through  alcohol, 
and  finally  treated  in  the  usual  way. 


Klein  investigated  cholera  in  India,  and  does  not  accept  Koch's 
conclusions.  With  regard  to  the  inoculation  experiments,  Klein 
believes  that  the  living  choleraic  comma -bacilli,  even  if  introduced 
in  large  numbers  into  the  small  intestine,  are  quite  innocuous, 
but  capable  of  great  multiplication  if  the  intestine  is  previously, 
from  some  cause  or  another,  diseased ;  the  chemical  products  of  the 
comma -bacilli  then  act  as  poisons  analogous  to  the  ptomaines 
obtained  from  other  putrefactive  bacteria.  The  observations  made 
by  Roy,  Brown,  and  Sherrington,  in  Spain,  tended  to  confirm 
Koch's  views.  Comma-bacilli  were  found  to  be  present,  in  some 
cases,  in  enormous  numbers,  and  the  frequency  of  their  occurrence 
led  these  observers  to  believe  that  they  must  bear  some  relation  to 

the  disease.  At  the  same  time, 
as  they  failed  to  find  them  in  all 
cases,  they  regarded  the  existence 
of  a  causal  relation  as  not  proven. 
They  failed  to  find  the  Naples 
bacterium,  or  a  small,  straight 
bacillus  noted  by  Klein ;  and 
they  drew  attention  to  certain 
FIG.  155. -COMMA-SHAPED  ORGANISMS  peculiar  mycelium-like  threads 

WITH    OTHER    BACTERIA    IN    SEWAGE- 
CONTAMINATED  WATER,  x  1200.  m  the  mucous  membrane  of  the 

intestines  ;  but  these  cannot  be 

considered  to  have  any  significance.  Methylene-blue  has  been 
employed  by  Koch  and  others,  including  the  author,  for  staining 
sections  of  the  intestine  from  cholera  cases,  and  had  they  been 
constantly  present,  it  is  hardly  possible  that  such  striking  objects 
could  have  been  overlooked.  Again,  we  must  bear  in  mind  that 
hypho-mycetous  fungi  occasionally  have  been  found  to  occur  sapro- 
phytically  in  the  intestinal  canal,  as  well  as  in  the  lungs,  external 
auditory  meatus,  and  elsewhere.  Cunningham,  of  Calcutta,  maintains 


ASIATIC   CHOLERA.  367 

that    Koch's   comma-bacilli   are   not   constantly    found ;    and   that 
the    comma -bacilli    obtained    from    typical   cholera   cases    show   a 
great   variation   in   cultivation,   and   cannot  be  distinguished  from 
comma-bacilli  from  other  sources. 
Cunningham      asserts      that 
comma-bacilli  resembling  Koch's 
are   found   in    the    intestine    in  * 

health.  Sternberg,  on  the  other 
hand,  made  a  number  of  examina- 
tions of  the  evacuations  of  yellow 

,  ,      ,,,       .    ,.       FIG.  156.— COMMA-BACILLI  OF  THE  MOUTH, 
fever  patients  and  healthy  mdi-  x  m  (VAN  ERMENGEM). 

viduals,   and  failed  to  find    any 

micro-organism  resembling  the  cholera  spirillum. 

Various  comma-bacilli  have  been  isolated  from  different  sources 
and  compared  with  Koch's  comma -bacillus.     Comma-bacilli  have  been 

found  in  the  mouth  by  Lewis  ; 

in  cholera  nostras  by  Finkler  and 
J^yf 

Prior ;    in    cheese    by    Deneke ; 

»'l)/(/r'       V.      ^  i11  nay  infusion  and  sewage   by 

Weibel ;  in  the  intestines  of  fowls 
by  Gamaleia,  and  in  water  by 
Sanarelli. 

Whether   the    comma-bacillus 

FIG.  157.— FINKLER'S  COMMA-BACILLI  ;  is  the  cause  of  cholera  or 
FROM  CHOLERA  NOSTRAS,  x  700  not?  its  detection  is  an  aid  in 
(FLUGGE).  ,.  .  T,.  ,  ,. 

diagnosis.      It    we    are    dealing 

with  a  case  alleged  to  be  one  of  Asiatic  cholera,  and  a  micro- 
organism is  found  in  the  intestinal  evacuations,  which  can  be 
differentiated  from  the  comma-bacillus  described  by  Finkler  in 
cholera  nostras,  and  identified 

with     the     comma-bacillus     de-  -^  . 

scribed  by  Koch,  we  are  justified  ^>  ^  \ 

in  regarding  the  case  as  one  of  "•»  *  p"*  ^v'Z.i'  •*    "* 

Asiatic  cholera.     But  we  cannot  ^~^*  •  ^  ^~* 

diagnose  Koch's  comma -bacillus,  '^^NfV*   f  ^X 

with  certainty,  unless  we  know 

the  source  of  the  culture.  The  FJG  m_DENEKE>s  COMMA-BACILLI, 
clinical  symptoms  of  cholera  in  FROM  CHEESE,  x  700  (FLUGGE). 

man,  and  especially  the  presence 

of  rice-water  stools,  must  be  taken  into  account,  together  with 
the  biological,  morphological,  and  chemical  characteristics  of 
the  bacilli  which  are  found  to  be  present.  There  are  several 


368  INFECTIVE    DISEASES. 

chemical  tests  which  can  be  applied  to  cultures.  According  to 
Frankel,  the  Bujwid-  Dunham  test  can  be  relied  upon  to  distin- 
guish Koch's  comma-bacillus  from  the  comma-bacillus  of  Finkler- 
Prior  (cholera  nostras),  and  from  those  found  by  Gamaleia.  The 
comma-bacilli  are  inoculated  in  broth  containing  peptone,  and, 
after  twelve  hours  in  the  incubator,  a  drop  of  strong  sulphuric 
acid  added  to  the  culture  will  produce  a  red  colour,  owing  to  the 
presence  of  indol.  A  test  which  distinguishes  Koch's  comma- 
bacillus  from  Finkler- Prior's  and  Deneke's  was  introduced  by  Cahen. 
A  solution  of  litmus  is  added  to  the  broth,  and  the  culture  placed 
in  the  incubator,  until  the  following  day  ;  in  the  case  of  Koch's 
commas,  the  colour  will  have  disappeared. 

Koch  points  out  that  in  the  bacteriological  diagnosis  of  cholera 
the  first  step  is  to  examine  the  mucus  in  the  evacuations,  or  in  the 
intestine  if  the  examination  is  made  after  death.  Cover-glass 
preparations  should  be  stained  with  dilute  Ziehl-Neelsen  solution. 
Cultures  are  next  made  in  peptone,  and  in  eight  hours  will  give  the 
indol  reaction.  In  twenty-four  hours  the  colonies  may  be  examined 
on  plate-cultivations.  The  peptone  cultures  are  prepared  by  adding 
a  trace  of  the  choleraic  evacuations,  or  of  mucus  containing  the 
bacilli,  to  a  sterilised  1  per  cent,  solution  of  peptone,  with  '5  to  1 
per  cent,  of  common  salt.  The  solution  must  be  alkaline,  and  the 
culture  is  placed  in  the  incubator  at  37°  C.  The  pathogenic  effects 
can  be  ascertained  by  diffusing  the  bacilli  from  an  agar-culture  in 
broth,  and  injecting  it  into  the  peritoneal  cavity. 

Toxic  Products. — Brieger  isolated  several  toxic  products  which  he 
had  found  in  association  with  putrefaction,  such  as  cadaverin  and 
putrescin  ;  but  there  were  also  present  two  new  toxic  substances,  one 
producing  cramps  and  muscular  tremors  in  inoculated  animals,  and 
the  other  lowering  the  temperature  and  depressing  the  action  of 
the  heart.  Later,  Brieger  in  conjunction  with  Frankel,  succeeded 
in  isolating  a  tox-albumin  from  pure  cultures.  Guinea-pigs  were 
killed  in  two  or  three  days,  but  rabbits  had  an  immunity.  Pfeiffer 
found  that  cultures  contained  a  poisonous  principle  which  proved 
fatal  to  guinea-pigs  in  extremely  minute  doses.  It  is  broken  up 
by  alcohol  and  by  boiling,  and  secondary  products  formed,  of  very 
much  mitigated  virulence.  Similar  toxic  products  were  obtained 
from  cultures  of  both  Finkler- Prior's  and  Metchnikoff's  commas. 

Protective  Inoculation — Haffkiiie  has  introduced  a  system  of 
protective  inoculation,  which  is  founded  on  the  principle  of  inducing 
the  formation  of  antitoxins,  or  defensive  proteids.  Comma-bacilli 
when  first  cultivated  from  a  cholera  patient  are  not  sufficiently 


ASIATIC   CHOLERA.  369 

virulent,  and  the  virulence  is  increased  by  cultivation  in  the 
peritoneal  cavities  of  a  succession  of  guinea-pigs.  This  successive 
cultivation  is  carried  on  until  a  virus  is  obtained  which  proves  fatal 
in  a  few  hours  when  inoculated  into  the  peritoneum.  A  culture 
from  the  peritoneum  is  obtained  on  an  agar  plate- cultivation,  and 
a  pure  sub-culture  on  agar  is  thoroughly  shaken  up  with  broth. 
This  constitutes  the  vaccinating  fluid.  It  may  be  used  as  a  living 
vaccine,  or  the  comma-bacilli  killed  by  the  addition  of  carbolic 
acid. 

Haffkine,  having  studied  the  pathological  and  physiological  effects 
on  some  sixty  persons,  mostly  scientists  interested  in  the  subject,  and 
finding  the  treatment  to  be  harmless,  transferred  his  operations  to 
localities  in  India  affected  by  cholera.  The  inhabitants  of  the 
northern  part  of  India  were  the  first  to  come  forward  and  submit 
themselves  to  the  inoculation.  In  the  course  of  the  first  year 
22,703  were  inoculated  in  the  North- West  Provinces  and  Oudh, 
and  in  the  Punjab.  All  classes  ofi  the  population  were  included. 
In  the  second  year  operations  were  carried  out  in  those  parts  of 
the  country  where  cholera  always  prevails,  and  where,  therefore,  the 
method  could  be  more  satisfactorily  tested. 

From  March  1894,  to  July  1895,  19,473  individuals  were 
inoculated  in  some  of  the  most  affected  localities. 

From  observations  made  at  Calcutta  by  Dr.  Simpson,  from  March 
1894  to  August  1895,  cholera  occurred  in  36  houses  containing 
inoculated  people.  There  were  521  inhabitants  in  the  infected 
houses,  of  whom  181  were  inoculated  from  1  to  459  days  before  the 
occurrence,  while  340  remained  uninoculated.  The  uninoculated  had 
45  cases  with  39  deaths  from  cholera ;  the  inoculated  had  4  deaths, 
1  occurring  451  days  after  the  first  inoculation,  and  3  others  from 
1  to  4  days  after  the  first  inoculation.  These  four  cases  had  not 
been  re- inoculated.  If  the  occurrences  in  inoculated  and  non-inocu- 
lated during  the  first  10  days  were  set  aside,  and  those  considered 
that  occurred  after  the  10  days  expired,  then,  according  to  Dr. 
Simpson,  the  proportion  of  cases  was  19 '2 7  and  that  of  deaths  17 '24 
times  smaller  in  the  inoculated  then  in  the  uninoculated. 

Cholera  broke  out  in  the  Gya  gaol,  and  inoculations  were  made 
after  6  cases,  with  5  deaths,  had  occurred.  During  the  stay  of  the 
prisoners  in  the  gaol,  there  were  209  uninoculated,  with  7  cases  and 
5  deaths,  and  211  inoculated,  with  5  cases  and  4  deaths. 

In  July  and  August  in  the  same  year  cholera  attacked  the  East 
Lancashire  Hegiment.  Out  of  773  men  there  were  133  inoculated 
and  640  uninoculated. 

24 


370 


INFECTIVE   DISEASES. 


The  occurrences  of  cases  and  deaths  were  : — 

In  640  uninoculated  120  cases  (18'75  %),  79  deaths  (12-34  %). 

In  133  inoculated  18  cases  (13-53  %),  13  deaths  (9'77  %). 

These  results  were,  it  is  said,  due  to  the  weakness  of  the  vaccines 
procurable  at  that  period  of  the  work,  and  to  the  small  doses  used. 

There  were  a  great  many  records  kept  of  the  results  of  inocula- 
tion of  coolies  on  tea  estates  in  different  localities.  After  a  summary 
of  the  results  Haffkine  concludes,  in  his  Report  to  the  Government 
of  India,  that,  in  his  opinion,  the  experimental  stage  was  not  yet 
in  so  advanced  a  condition  as  to  be  completely  closed;  but  that 
the  observations  made  and  records  collected  justified  steps  being 
taken  to  give  the  inoculations  a  more 
extended  trial. 

CHOLERA  NOSTRAS. 

Cholera  nostras,  English  cholera,  or 
English  dysentery,  produces  an  inflamma- 
tion of  the  mucous  membrane  of  the 
bowels  with  croupous  exudation.  The 
large  intestine  is  commonly  affected,  and 
the  mucous  membrane  may  be  covered 
with  small  superficial  ulcers.  The  disease 
is  associated  with  severe  diarrhoea. 

Finkler  and  Prior  obtained  a  comma- 
bacillus  from  the  evacuations,  which  they 
believed  to  be  identical  with  the  comma- 
bacillus  found  by  Koch  in  Asiatic  cholera. 
Koch  pointed  out  that  there  were  marked 
differences  in  the  biological  character  of 
the  two  micro-organisms. 

Spirillum  Finkler-Prior  (Comma- 
bacillus  in  Cholera  nostras). — Curved  rods, 
thicker  than  the  comma-bacillus  of  Koch, 
and  spirilla.  The  colonies  on  plate- 
cultivations  are  very  much  larger  than 

those  of  the  comma -bacillus  of  Koch  of  the  same  age.  They  have  a 
very  faint  yellowish-brown  tinge,  a  well-defined  border,  and  a  distinctly 
granular  appearance.  They  liquefy  nutrient  gelatine  very  rapidly, 
so  that  the  first  plate  of  a  series  is,  as  a  rule,  completely  liquefied  on 
the  day  following  inoculation,  and  the  second  plate  in  two  or  three 
days  more.  In  a  test-tube  cultivation  in  nutrient  gelatine  the 
appearances  are  especially  characteristic  :  the  gelatine  is  very  rapidly 


FIG.  159.— PURE-CULTIVATION 
OF  THE  SPIRILLUM  ^INKLER- 
PRIOR,  IN  NUTRIENT  GELA- 
TINE. In  thirty-six  hours. 


CHOLERAIC   DIARRHCEA   FROM   MEAT   POISONING.  371 

liquefied  along  the  whole  track  of  the  needle,  so  that  the  cultivation 
resembles  a  conical  sack,  or  the  finger  of  a  glove  turned  inside  out. 
On  a  sloping  surface  of  nutrient  agar-agar  a  white  moist  layer  forms 
very  quickly.  On  potato  they  grow  at  the  ordinary  temperature 
of  the  air,  producing  a  brownish  layer  and  corrosion  of  the  surface 
of  the  potato.  They  have  been  shown  to  be  pathogenic. 

CHOLERAIC  DIARRHCEA  FROM  MEAT  POISONING. 

There  are  two  varieties  of  choleraic  diarrhoea  from  meat  poisoning, 
and  both  are  associated  with  vomiting,  diarrhoea,  pain  in  the  abdo- 
men, in  severe  cases  followed  by  suppression  of  urine,  collapse,  and 
death.  These  conditions  are  brought  about  by  poisonous  foods,  and 
include  those  cases  of  poisoning  by  tinned  meats,  pork  pies,  hams, 
cheese,  sardines,  and  other  articles  of  food  improperly  prepared.  In 
most  cases  putrefaction  has  taken  place,  owing  to  the  action  of 
various  bacteria.  Associated  with  their  growth  we  find  highly 
poisonous  substances,  but  no  bacteria  are  found  in  the  body  in  these 
cases.  They  are  all  due  to  chemical  poisoning  ;  but  Klein  has  also 
described  cases  of  poisoning  due  to  the  growth  of  bacteria  without 
the  presence  of  putrefaction.  The  latter  were  of  the  nature  of  an 
infectious  disease.  In  the  Welbeck  poisoning  cases,  described  by 
Ballard,  the  poisonous  hams  contained  a  short  bacillus,  which  was 
also  found  in  the  kidney  and  spleen  in  the  fatal  cases  in  man.  In 
the  Carlisle  epidemic,  which  was  due  to  poisonous  pork  pies,  the  pork 
and  gravy  stock  proved  fatal  to  mice,  and  from  the  infected  mice 
a  bacillus  was  cultivated,  which,  administered  to  mice  by  feeding  or 
subcutaneous  inoculation,  produced  enteritis,  diarrhoea,  and  congestion 
of  the  lungs. 

Gartner  cultivated  Bacillus  enteritidis  from  the  spleen  in  a 
fatal  case  of  meat  poisoning.  Gaffky  obtained  a  similar  bacillus  in 
cases  of  gastro-enteritis,  following  the  consumption  of  meat  and 
sausages,  which  had  been  made  of  horseflesh. 

Bacillus  of  Choleraic  Diarrhoea  from  Meat-poisoning 
(Klein). — Rods  from  3  to  9  /A  in  langth,  1*3  /x  wide,  rounded  at  their 
extremities,  singly  or  in  chains  of  two.  Spore- formation  occurs, 
the  spores  being  1  //,  thick,  ovtil,  and  situated  in  the  centre  or  at 
the  end  of  the  rod. 

Feeding  mice  with  the  bacilli  and  inoculation  produced  positive 
results.  At  the  autopsy,  pneumonia,  peritonitis,  pleuritis,  enlargement 
of  the  liver  and  spleen,  and  haemorrhages  were  observed,  and  bacilli 
were  present  in  the  blood  and  exudations  of  these  animals.  They 


372 


INFECTIVE    DISEASES. 


occurred  in  the  blood  and  juices,  and  especially  in  the  glomeruli  of  the 
kidneys,  of  several  fatal  cases  of  choleraic  diarrhoea. 

Bacillus  enteritidis  (Gartner). — Short  rods  in  pairs,  and  short 
chains.  They  are  motile  ;  spore-formation  not  observed.  Colonies 
are  granular,  and  old  colonies  at  the  margin  have  an  appearance  of 


FIG.  160.— TROPICAL  DYSENTERY.  Mucous  membrane  of  large  intestine  some 
months  after  an  acute  attack  :  a,a,  representing  remains  of  mucosa  ;  6,6,  inter- 
vening parts  corresponding  to  the  muscularis  (HAMILTON). 

powdered  glass.  On  the  surface  of  gelatine  a  thick  greyish-white 
film  develops,  which  in  time  becomes  wrinkled.  In  the  depth  of 
gelatine  a  white  filament  forms.  The  gelatine  is  riot  liquefied.  On 
agar  the  film  is  slightly  yellowish.  On  potato  it  is  similar  in  colour, 
moist  and  shining.  On  blood  serum  it  is  very  similar.  Mice  fed 


CHOLERAIC    DIARRHCEA   IN   FOWLS.  373 

with  the  bacilli  die  in  one  or  two  days.  Subcutaneous  injection  is 
fatal  in  guinea-pigs  and  rabbits  in  from  two  to  five  days.  Dogs, 
cats,  and  fowls  are  immune. 

The  bacilli  were  obtained  from  a  cow  suffering  from  a  disease 
associated  with  diarrhoea,  and  from  the  spleen  of  a  man  who  died 
twelve  hours  after  partaking  of  the  flesh  of  this  animal. 

DYSENTERY. 

Dysentery  is  a  disease  of  tropical  climates  associated  with  in- 
flammation and  ulceration  of  the  large  intestine  (Fig.  160).  At  first 
the  discharge  from  the  bowel  is  a  whitish  or  brownish  mucus, 
which  soon  becomes  blood-stained;  later  the  evacuations  become 
thin  and  watery,  with  altered  blood  clots,  fragments  of  mucous 
membrane,  and  pieces  of  false  membrane  ;  and  in  some  cases  they 
become  purulent.  The  virus  is  believed  to  be  in  the  intestinal 
discharges,  which  by  contaminating  water  or  soil  may  give  rise  to 
other  cases. 

Micrococci  have  been  found  in  dysentery,  but  the  micro-organism 
which  has  received  most  attention  is  a  protozoon,  the  Amceba  coli, 
which  will  be  described  in  another  chapter. 

CHOLERAIC  DIARRHOEA  IN  FOWLS. 

Choleraic  diarrhoea  in  fowls,  or  gastro-enteritis  cholerica,  is  an 
infectious  disease  of  fowls,  occurring  in  Russia  during  the  summer. 
The  disease  is  very  like  fowl-cholera.  The  birds  are  sleepy,  and 
suffer  from  diarrhoea,  but  the  temperature  is  not  raised,  as  in 
fowl-cholera.  After  death  there  is  usually  an  abundance  of  greyish 
liquid  in  the  small  intestine,  which  is  stained  with  blood.  It  was 
investigated  by  Gamaleia,  who  found  a  comma-bacillus,  to  which  he 
gives  the  name  Vibrio  Metchnikovi. 

Spirillum  of  Fowl- enteritis  (Vibrio  Metchnikovi). — Curved 
rods  and  spirilla;  thicker,  shorter,  and  more  curved  than  Koch's 
commas.  They  are  motile,  and  possess  a  single  flagellum  at  one  end. 
They  stain  with  the  usual  dyes.  Spore-formation  doubtful.  In  plate- 
cultivations  minute  white  colonies  appear  in  from  twelve  to  sixteen 
hours,  and  the  gelatine  is  liquefied.  The  colonies  in  about  three 
days  resemble  those  of  both  Finkler- Prior's  and  Koch's  comma-bacilli, 
some  colonies  being  more  like  the  one  kind,  and  some  like  the  other. 
In  the  depth  of  gelatine  the  growth  is  very  much  like  that  of  Koch's 
comma-bacillus,  possessing  the  characteristic  air-bubble  appearance. 


374  INFECTIVE    DISEASES. 

On  agar  a  slightly  yellowish  growth  is  obtained,  resembling  that 
of  Koch's  commas  ;  on  potato  a  yellowish -brown  or  chocolate  layer 
develops  after  incubation  at  the  temperature  of  the  blood,  very 
similar  to  cultures  from  Asiatic  cholera.  Broth  becomes  turbid,  and 
a  wrinkled  film  forms  on  the  surface  ;  the  addition  of  sulphuric  acid 
gives  the  indol  test.  The  spirilla  grow  in  milk,  and  coagulate  it ; 
the  milk  becoming  strongly  acid,  and  the  casein  being  precipitated. 
They  are  pathogenic  in  chickens,  pigeons,  and  guinea-pigs.  Pigeons 
die  in  about  twelve  hours  after  a  subcutaneous  injection  ;  and  the 
spirilla  are  found  abundantly  in  blood  from  the  heart.  Guinea-pigs 
die  from  acute  septicaemia  in  about  twenty-four  hours.  The  spirilla 
are  found  in  the  blood  and  internal  organs.  Inoculation  of  pigeons 
and  guinea-pigs  with  sterilised  cultures  will  produce  immunity. 


CHAPTER    XXVIII. 

TUBERCULOSIS. 

TUBERCULOSIS  is  a  communicable  disease  of  man  and  animals,  charac- 
terised by  the  formation  of  new  growths  associated  with  the  presence 
of  the  tubercle  bacillus.  Yon  Bayle,  in  1810,  was  the  first  to  describe 
little  growths  like  millet  seeds,  which  were  considered  to  be  character- 
istic of  consumption  or  phthisis.  Laennec,  in  1834,  attached  much 
more  importance  to  the  existence  of  caseous  matter  and  classified 
miliary  tubercle,  crude  tubercle,  granular  tubercle,  and  encysted 
tubercle,  as  varieties  of  tuberculosis.  Virchow  would  not  accept 
all  these  varieties  as  tubercular,  and  only  regarded  those  conditions 
associated  with  the  presence  of  miliary  tubercles  as  genuinely 
tubercular.  Laennec's  so-called  crude  tubercle,  for  example,  was 
simply  due  to  pneumonic  caseation.  Yillemin  threw  entirely  fresh 
light  upon  this  controversy  by  proving  that  tuberculosis  was  a 
communicable  disease.  Rabbits  and  guinea-pigs,  inoculated  with 
tubercular  sputum  or  caseous  tubercle,  developed  miliary  tubercle  in 
a  few  weeks.  Sanderson  confirmed  these  experiments,  and  pointed 
out  that  foreign  bodies  would  produce  experimental  tuberculosis 
in  rabbits.  Cohnheim  also  confirmed  the  experiments  of  Villemin, 
and  maintained  that  tuberculosis  was  a  specific  inoculable  disease, 
and,  therefore,  everything  was  tubercular  which,  on  inoculation, 
produced  tuberculosis.  Koch,  in  1882,  announced  the  discovery  of 
the  tubercle  bacillus,  and  expressed  the  opinion  that  without  the 
tubercle  bacillus  there  could  be  no  tuberculosis.  Tubercle  was 
defined  as  tissue  containing  the  tubercle  bacillus,  whatever  might 
be  the  clinical  manifestations  of  the  case,  or  the  microscopical  and 
naked-eye  appearances  of  the  diseased  parts. 

A  tubercle  is  a  small  growth  about  the  size  of  a  millet  seed.  In 
the  early  stage  it  is  circular,  hard,  grey  in  colour,  and  lustrous ;  but 
when  it  undergoes  necrosis  and  caseation  it  becomes  soft  and  yellowish. 
In  the  very  early  stage  it  consists  of  a  little  collection  of  round 

375 


376 


INFECTIVE   DISEASES. 


cells,  in  which  it  is  possible,  though  often  with  extreme  difficulty, 
to  demonstrate  the  tubercle  bacillus.  The  cells  originate  in  the 
proliferation  of  endothelial  connective  tissue  and  white  blood  cells. 
Later  on,  large  oval  or  circular  multi- nucleated  cells,  or  giant  cells, 
make  their  appearance.  The  tubercle  bacilli  are  only  occasionally 
found  in  the  interior  of  human  giant  cells,  whereas  in  the  lower 
animals,  in  equine  and  bovine  tuberculosis  more  especially,  the  bacilli 
are  often  present  in  great  numbers,  and  very  commonly  in  the  form 
of  conspicuous  rings,  visible  under  a  low  power  of  the  microscope. 


FIG.  161.— TUBERCLE  OF  THE  LUNG  IN  A  VERY  EARLY  STAGE,  x  400  :  a,  An  alveolar 
wall ;  b,  blood-corpuscles  in  capillaries  of  the  same ;  c,  blood-corpuscles 
extra vasated  into  the  alveolar  cavities;  d,  alveolar  capillaries  filled  with 
blood-corpuscles  carried  forward  by  the  tubercle  which  is  growing  into  the 
alveolar  cavity ;  e,  large  endothelium-like  cells,  of  which  the  tubercle  in  this 
stage  is  mainly  composed  ;  /,  portion  of  a  branch  of  the  pulmonary  artery 
injected  (HAMILTON). 

Whether  the  absence  of  blood-vessels  or  the  action  of  the  bacillus  is 
the  main  factor  in  producing  caseation,  is  an  open  question.  When 
suppuration  follows  caseation,  as  commonly  happens  in  tuberculosis 
of  the  lungs  in  man,  and  in  experimental  tuberculosis  in  animals,  an 
abscess  forms.  In  cattle  there  is  a  remarkable  tendency  to  the 
formation  of  calcareous  deposit  in  the  caseous  masses. 

The  tubercle  may  not  degenerate  and  die,  but  live  and  develop. 


TUBERCULOSIS. 


377 


The  giant  cells,  which  are  more  or  less  central,  have  been  described 
as  sending  off  processes,  which,  by  dividing  and   subdividing,  and 


FIG.  162. — PRIMARY  TUBERCLE  OF  LUNG  TWO  TO  THREE  WEEKS  OLD,  x  50: 
a,  Portion  of  wall  of  a  branch  of  the  pulmonary  artery  ;  b,b,  giant  cells  with 
concentric  arrangement  of  fibrous  tissue ;  c,  centre  of  tubercle  beginning  to 
caseate ;  d,  small  branch  of  pulmonary  artery  seen  on  transverse  section ; 
e,  injected  capillaries  of  the  alveolar  walls  (HAMILTON). 


interlacing,  form  a  reticnlum,  or 
meshwork.  Towards  the  periphery 
of  the  tubercle  the  reticulum  may 
become  arranged  in  the  form  of  a 
capsule  as  the  age  of  the  tubercle 
advances,  and  the  reticular  giant 
cell  becomes  eventually  converted 
into  fibrous  tissue.  The  bacillus  has 
disappeared,  and  the  tubercle  has 
healed. 

Giant  cells  cannot  be  relied  upon 
to  indicate  tuberculosis.  They  are 
not  always  present  in  tubercu- 
losis, and  they  are  not  peculiar  to 
tubercle,  being  found,  for  example, 
in  actinomycosis.  The  only  certain 
indication  of  tuberculosis  is  the  pre- 
sence of  the  tubercle  bacillus,  which 


FIG.  163.— LA  RGB  OVAL  GIANT  CELL 
FROM  TUBERCLE  OF  LUNG  x  300 : 
a,  Granular  centre;  b,  nucleated 
periphery  forming  a  mantle-like 
sheath ;  c,  processes  from  the 
same. 


378  INFECTIVE    DISEASES. 

can  be  revealed  either  by  microscopical  examination  of  the  suspected 
tissue,  or  after  inoculation  in  guinea-pigs. 

Bacillus  Tuberculosis  (Koch). — Rods,  2  to  4  //,  and  occa- 
sionally 8  /A  long,  very  thin,  and  rounded  at  the  ends.  They  are 
straight  or  curved,  and  frequently  beaded,  and  occur  singly,  in  pairs, 
or  in  bundles ;  there  are  also  involution  forms  and  short  branched 
threads.  Spore-formation  is  observed  in  old  cultures.  They  are 
non- motile.  Tn  the  interior  of  giant  cells  they  are  often  accompanied 
by  grains  which  exhibit  the  same  colour  reaction. 

The  bacilli  in  tissue  sections  of  bovine  tuberculosis  are  shorter 
and  less  granular  than  those  in  human  tubercular  sputum,  but  in 
milk  they  are  quite  as  long,  and  even  longer,  and  very  distinctly 
granular  or  beaded,  and  are  thus  brought  much  closer,  morpho- 
logically, to  the  bacilli  in  human  sputum.  Speaking  generally, 
however,  the  average  length  of  the  human  bacilli  is  greater  than 
the  average  length  of  the  bacilli  in  cow's  milk,  but  the  longest  of 
the  bovine  bacilli  cannot  be  distinguished  in  length  from  the  longest 
human  bacilli.  There  are,  however,  exceptional  cases,  for  in  some 
preparations  of  pus  from  human  lungs  the  bacilli  are  remarkable, 
not  only  for  their  thinness,  and  their  uniformly  beaded  character, 
but  more  particularly  for  their  extraordinary  length.  They  should 
be  compared  with  other  preparations,  in  which  the  bacilli,  though  in 
human  sputum,  are  sometimes  much  more  distinctly  rod-shaped, 
much  shorter  and  thicker,  with  complete  absence  of  any  beaded 
appearance.  Neither  length  nor  granularity  is  a  characteristic 
sufficient  to  denote  any  specific  difference  between  human  and  bovine 
bacilli.  The  author  has  examined  minutely  the  bacilli  in  tuberculosis 
of  other  animals,  such  as  the  horse,  pig,  and  cat ;  and  of  birds — the 
fowl,  guinea-fowl,  pheasant,  and  ostrich.  Here,  again,  minute 
morphological  differences  can  be  observed.  For  example,  in  many 
cases  in  fowls  the  bacilli  are  conspicuously  short  and  straight.  In 
the  liver  and  lungs  of  an  ostrich,  packets  of  short  rod-forms  are 
found,  while  in  other  parts  of  the  same  sections  the  bacilli  attain 
a  very  great  length.  Many  of  the  long,  sinuous  forms  exhibit  a 
peculiar  terminal  enlargement.  There  are  also  short  rods  with  a 
similar  appearance,  and  free  ovoid  bodies,  singly  and  in  groups,  which, 
from  their  connection  with  the  bacilli,  and  their  sharply  defined 
outline  in  the  free  state,  are  similar  to  spores  in  old  cultures. 

Thus,  morphological  differences  are  found  under  different  circum- 
stances, and  within  limits  the  morphology  of  the  tubercle  bacillus 
varies  with  its  environment. 

Koch   first  succeeded  in  cultivating  the  bacillus  by  employing 


DESCRIPTION    OF    PLATE    XI. 
Bacillus  tuberculosis. 

The  figures  in  this  plate  represent  the  bacilli  of  tuberculosis  in 
different  animals,  examined  under  the  same  conditions  of  amplifica- 
tion and  illumination.  x  1200.  Lamp-light  illumination. 

FlG.  1. — Bacilli  in  pus  from  the  wall  of  a  human  tubenmlar  cavity.  In 
this  specimen  the  bacilli  are  shorter  than  those  in  tubercular  sputum, 
and  are  very  markedly  beaded. 

FIG.  2. — Bacilli  in  pus  from  a  tubercular  cavity  from  another  case  in  man. 
They  are  present  in  the  preparation  in  enormous  numbers.  The  proto- 
plasm occupies  almost  the  whole  of  the  sheath,  and  the  bacilli  are 
strikingly  thin  and  long. 

FlG.  3. — Bacilli  in  sputum  from  an  advanced  case  of  phthisis,  showing 
the  ordinary  appearance  of  bacilli  in  sputum  ;  some  beaded,  others 
stained  in  their  entirety ;  occurring  both  singly  and  in  pairs,  and 
in  groups  resembling  Chinese  letters. 

FlG.  4. — Bacilli  in  a  section  from  the  lung  in  a  case  of  tuberculosis  in  man. 
The  bacilli  in  human  tuberculosis  are  found  in,  and  between,  the  tissue 
cells  ;  and  sometimes,  as  in  equine  and  bovine  tuberculosis,  in  the 
interior  of  giant  cells,  but  not  so  commonly. 

FIG.  5. — From  a  cover-glass  preparation  of  the  deposit  in  a  sample  of  milk 
from  a  tubercular  cow.  The  bacilli  were  longer  than  the  average 
length  of  bacilli  in  bovine  tissue  sections,  and  many  were  markedly 
beaded. 

FIG.  6. — From  a  section  of  the  brain  in  a  case  of  tubercular  meningitis  in  a 
calf,  showing  a  giant  cell  containing  bacilli  with  the  characters  usually 
found  in  sections  of  bovine  tuberculosis. 

FIG.  7. — From  a  section  of  the  liver  of  a  pig  with  tubercle  bacilli  at  the 
margin  of  a  caseous  nodule. 

FIG.  8. — From  a  cover-glass  preparation  of  a  crushed  caseous  mesenteric 
gland  from  a  rabbit  infected  by  ingestiori  of  milk  from  a  cow  with 
tuberculosis  of  the  udder. 

FlG.  9.— From  a  section  of  lung  in  a  case  of  equine  tuberculosis,  showing  a 
giant  cell  crowded  with  tubercle  bacilli. 

FIG.  10. — From  a  section  of  lung  from  a  case  of  tuberculosis  in  the  cat,  with 
very  numerous  tubercle  bacilli. 

FIG.  11. — From  a  cover-glass  preparation  of  a  crushed  caseous  nodule  from 
the  liver  of  a  fowl,  with  masses  of  bacilli.  These  are  for  the  most  part 
short,  straight  rods ;  but  other  forms,  varying  from  long  rods  to  mere 
granules,  are  also  found. 

FIG.  12. — From  sections  of  the  liver  and  of  the  lung  in  a  case  of  tubercu- 
losis of  a  Khea.  Isolated  bacilli  are  found,  as  well  as  bacilli  packed  in 
large  cells,  colonies  of  sinuous  bacilli,  and  very  long  forms  with  terminal 
spore-like  bodies  and  free  oval  grains. 

The  preparations  from  which  these  figures  were  drawn  were  all 
stained  by  the  Ziehl-Neelsen  method,  with  the  exception  of  the 
first,  which  was  stained  by  Ehrlich's  method. 


Plate  XL. 


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,'. 


A 


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"Fit*  3 


Fig  5. 


v- 


Pig  6 


X 


Pig  7. 


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-7      \    a  ' 

x^  ^^ 


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' 


BACILLUS  TUBERCULOSIS. 


•vTlBKA/fp1 

OF  THB 

"UNIVERSITY 

Or 


TUBERCULOSIS.  379 

blood  serum.  Solid  blood  serum,  with  or  without  the  addition  of 
gelatine,  was  employed,  and  the  cultures  incubated  at  37°  C.  The 
growth  takes  place  very  slowly,  and  only  between  the  temperatures 
of  30°  C.  and  41°  C.  In  about  eight  or  ten  days  the  growth  appears 
as  little  whitish  or  yellowish  scales  and  grains. 

The  bacillus  can  also  be  cultivated  in  a  glass  capsule,  on  blood 
serum,  and  the  appearances  of  the  growth  studied  under  the 
microscope.  The  scales  or  pellicles  were  described  by  Koch  as  made 
up  of  colonies  of  a  perfectly  characteristic  appearance,  which  may 
be  still  further  studied  by  making  a  cover-glass  impression.  They 
are  then  seen  to  be  composed  of  bacilli,  arranged  more  or  less 
with  their  long  axis  corresponding  with  that  of  the  colony  itself, 
and  with  an  appreciable  interval  between  the  individual  bacilli. 
The  colonies  themselves  appear  as  fine  curved  lines,  the  smallest 
being  mostly  S~snaPe(i-  Longer  colonies  have  serpentine  twistings 
and  bendings,  which  often 
recall  the  curves  of  fancy 
lettering.  The  ends  of  the 
lines  run  to  sharp  points, 
but  the  middle  of  the 
growth  is  spindle -formed. 
The  youngest  colonies  are 
extremely  delicate  and 
narrow,  but  the  older 
colonies  increase  in  size, 

are    thicker    across,     and,      FlG-    164.  -  BACILLUS    TUBERCULOSIS,    FROM 

TUBERCULAR  SPUTUM,  x  2500.    From  Photo- 
blending  with  each  other,          graphs 

gradually     obliterate     the 

characteristic  appearances ;  a  lamellated  growth  results,  which 
increases,  and  gives  the  appearance  to  the  naked  eye  of  the  scale 
or  pellicle  already  described.  The  blood  serum  is  not  liquefied 
unless  putrefactive  bacteria  contaminate  the  culture.  A  fresh  tube 
can  be  inoculated  with  one  of  the  little  scales,  and  a  new  generation 
started.  The  scales  gradually  increase  in  size,  and  consist  entirely 
of  bacilli.  In  about  three  to  four  weeks  the  cultivation  ceases  to 
increase,  and  it  is  then  necessary  to  inoculate  a  fresh  tube. 

In  liquid  blood  serum  a  film  forms  on  the  surface  of  the  liquid, 
and  is  easily  broken  by  agitation.  In  neutralised  broth  there  is 
very  little  indication  of  success.  When  a  triturated  culture  is  added 
to  the  broth,  a  granular,  sandy,  whitish  deposit  collects  at  the 
bottom  of  the  vessel,  with  indications  of  an  increase  in  amount. 
Koch  also  tried  nutrient  agar-agar,  which  did  not  prove  to  be  at 


380 


INFECTIVE    DISEASES. 


BACILLUS    TUBERCULOSIS. 


FIG.  165. — PURE-CULTIVATIONS  ON  GLYCERINE-AGAR  FROM  HUMAN  TUBERCULAR 
SPUTUM,  a,  After  six  months'  growth.  (Fifth  sub-culture.)  b  and  c,  After 
ten  months'  growth.  (Fourth  sub-cultures.) 


TUBERCULOSIS. 


381 


all  a  favourable  medium.      Some   increase   took   place,   but   there 
was  no  continuous  growth  over  the  inoculated  area. 

Glycerine  Agar-agar. — Nocard  and  Roux  were  among  those  who 
worked  at  the  subject  and  confirmed  Koch's 
observations,  Nocard  attempted  to  get 
cultures  of  avian  tuberculosis  on  blood  serum 
to  which  peptone,  salt,  and  cane  sugar  had 
been  added.  The  results  were  more  success- 
ful than  with  ordinary  serum.  But  he 
encountered  a  difficulty  in  the  rapid  drying 
of  the  surface  of  the  medium,  which  rendered 
the  tubes  unfit  for  use.  It  occurred  to 
Nocard  and  Roux  to  obviate  this  by  the 
addition  of  a  hygroscopic  agent,  and  accord- 
ingly they  added  sterilised  glycerine.  The 
result,  which  far  exceeded  their  expectation, 
evidently  was  not  solely  attributable  to  the 
prevention  of  desiccation.  Following  up 
their  discovery,  and 
being  anxious  to  find 
a  medium  more  easily 
prepared  than  blood 
serum,  they  added 

6  to  8  per  cent,  of  glycerine  to  ordinary 
nutrient  agar-agar.  The  bacillus  grew  so 
abundantly  in  this  mixture  that  a  culture 
in  fifteen  days  equalled  in  extent  a  culture 
on  blood  serum  which  was  several  weeks  old. 
The  bacillus  was  found  to  grow  abundantly 
in  veal  broth,  to  which  glycerine  had  been 
added  in  the  proportion  of  5  per  cent.,  the 
bottom  of  the  flask  being  covered  in  about 
three  weeks  with  a  flocculent  deposit,  having 
some  resemblance  to  anthrax  cultivations 
in  liquid  media.  In  beef  broth,  chicken 
broth,  and  in  Cohn's  liquid,  cultures  were 
obtained  after  the  addition  of  glycerine. 

Description  of  Cultivations  on   Glycerine 
Agar-agar. — The  cultivations  on  the  sloping 
surface  of  obliquely  solidified  glycerine  agar- 
agar  begin  to  appear  in  from  four  to  six  days  as  very  minute  white 
colonies.     These  steadily  increase  in  size,  and  either  look  moist  and 


FIG.  166.— PURE-CULTI- 
VATION IN  GLYCERINE 
AGAR-AGAR,  after  ten 
months'  growth. 


FIG.  167. -PURE-CULTI- 
VATION IN  GLYCERINE 
AGAR-AGAR, — A  SUB- 
CULTURE FROM  A  PURE- 
CULTURE  IN  GLYCERINE- 
MILK.  In  two  months. 


382  INFECTIVE    DISEASES. 

smooth,  or,  even  at  this  early  stage,  appear  dry  and  crinkled. 
According  to  the  number  of  bacilli  inoculated,  the  colonies  will 
either  remain  isolated  or  coalesce  and  form  a  more  or  less  continuous 
film.  If  the  nutrient  agar-agar  has  only  recently  been  prepared, 
there  is  usually  a  quantity  of  liquid  present,  and  the  bacillus  forms 
a  white  coating  over  the  inoculated  area  and  beyond  it.  The 
appearances  are  much  more  characteristic  when  this  medium  is, 
comparatively  speaking,  dry.  A  semi-transparent  membranous 
growth  develops,  thickens,  and  assumes  a  characteristic  lichenous 
appearance.  Such  a  culture,  examined  with  a  pocket  lens,  resembles 
a  model  in  wax  in  miniature  of  the  folds  of  the  gastric  mucous 
membrane.  In  about  six  weeks  to  two  months  the  culture  has  fully 
developed.  In  old  cultures,  especially  when  the  individual  colonies 
remain  isolated,  the  appearance  is  very  characteristic.  Some  cul- 
tures in  appearance  closely  resemble  cultivations  on  blood  serum. 
The  consistency  of  the  growth  depends  upon  the  character  of  the 
soil  and  the  age  of  the  culture.  When  the  medium  is  moist  the 
growth  is  moist  and  viscous,  but  more  often  it  is  distinctly  tallowy, 
and  in  old  and  dry  cultures  scaly  and  friable. 

Cultivations  in  Glycerine  Broth. — In  a  few  days  minute  flakes  are 
visible,  which  steadily  increase  in  size,  and  subside  to  the  bottom,  of 
the  flask,  forming  in  time  a  very  copious  deposit.  On  shaking  the 
flask,  this  deposit,  which  is  extremely  tenacious,  rises  in  stringy 
masses,  and  gives  an  appearance  which  is  more  or  less  character- 
istic. If  the  flask  is  left  undisturbed,  a  delicate  veil-like  film  forms 
over  the  surface,  which  can  be  readily  broken  up  by  gentle  agitation, 
forming  flakes  which  gradually  sink  in  the  liquid.  If  undisturbed 
for  several  weeks  this  film  increases  in  thickness,  is  irregularly 
fissured,  and  has  more  the  appearance  of  masses  of  tallow  floating 
on  the  surface.  The  growth  also  may  be  seen  to  extend  up  the  side 
of  the  flask  above  the  liquid.  Pasteur  or  Erlenmeyer  flasks  can  be 
employed  for  these  cultures.  Solidified  egg-albumin  added  to  the 
glycerine  broth  seems  to  increase  the  amount  of  growth,  which 
clings  to  the  albumin,  and  waves  to  and  fro  in  the  liquid  when  the 
flask  is  gently  shaken.  The  author  has  confirmed  the  observation 
of  Nbcard  and  Roux,  that  sub-cultures  from  glycerine  agar-agar,  or 
from  glycerine  broth,  will  give  cultures  in  ordinary  broth  without 
glycerine.  Ordinary  broth  with  egg-albumin,  and  without  glycerine, 
will  also  give  a  good  growth  when  inoculated  from  previous  sub 
cultures,  although  the  attempt  to  produce  primary  cultures  in  these 
media  has  hitherto  failed. 

Cultivations   in  Glycerine- Milk,  and  other   Media. — In  milk  the 


TUBERCULOSIS.  383 

author  found  there  was  only  a  slight  increase  in  the  number  of  bacilli 
inoculated,  but  milk  with  glycerine  in  the  proportion  of  5  per  cent, 
forms  a  more  favourable  medium.  The  author  has  also  cultivated  the 
bacillus  on  sterilised  urine  and  glycerine,  and  ordinary  nutrient 
gelatine  with  5  per  cent,  of  glycerine.  On  potato  the  growth  of 
the  bacillus  is  extremely  slow.  Beevor  succeeded  in  obtaining 
cultures  at  the  ordinary  temperature  of  the  room. 

Examination  of  Cultivations. — To  examine  the  bacilli  in  these 
various  preparations  the  author  prefers  to  use  Neelsen's  method, 
floating  the  cover-glasses  for  from  five  to  ten  minutes  on  warm 
carbolised  fuchsine,  and  passing  them  through  dilute  sulphuric  acid. 
In  some  cultures  the  bacilli  are  shorter  and  thicker  than  is  commonly 
observed  in  human  sputum,  and  they  are  for  the  most  part  without 
the  beaded  appearance.  In  old  cultures  on  glycerine  agar-agar  the 
number  of  granular  or  beaded  bacilli  increases,  and  there  are  also 
numerous  peculiar  forms.  There  are  bacilli,  sometimes  two  or 
three  times  the  length  of  an  ordinary  bacillus,  provided  with  a 
club-shaped  enlargement  at  one  or  both  extremities,  and  in  rare 
cases  with  lateral  branches.  They  are  no  doubt  identical  with  the 
bacilli  with  swollen  extremities  and  the  branched  forms  observed 
by  Nocard  and  Roux. 

In  milk  the  appearance  is  very  striking,  many  bacilli  attaining 
in  old  cultures  a  great  length,  and  all  are  more  uniformly  beaded 
than  in  any  other  cultivations.  Staining  preparations  by  the  method 
of  Grain  in  all  cases  exaggerate  this  appearance. 

The  important  part  played  by  the  environment  is  shown  by  the 
morphological  differences  observed  in  artificial  cultivation  under 
varying  conditions,  and  by  the  fact  that  by  successive  cultivation 
the  bacillus  can  be  educated  to  grow  upon  a  medium  which  is  un- 
suitable for  obtaining  primary  cultures. 

Impression  preparations  of  the  growth  of  the  bacillus  on  the 
surface  of  glycerine  agar-agar  in  capsules  show  a  tendency  to  the 
formation  of  serpentine  colonies,  composed  of  bundles  of  more  or 
less  parallel  bacilli. 

Spore-formation. — In  old  cultivations  true  spore-formation  can 
readily  be  observed,  both  in  stained  and  unstained  preparations.  In 
the  latter  case  they  are  recognised  in  the  form  of  one  or  two  highly 
refractive  bodies  in  individual  bacilli. 

Toxic  Products  of  Cultures. — The  poisonous  substances  found  in 
cultures,  and  the  composition  and  use  of  tuberculin,  have  already 
been  described  (p.  43). 

Inoculation  Experiments. — A  relatively  small  portion  of  a  culti- 


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^w.    r>  _    • 


384  INFECTIVE    DISEASES. 

vation  inoculated  into  the  subcutaneous  tissue,  into  the  peritoneal 
or  pleural  cavities,  into  the  anterior  chamber  of  the  eye,  or  directly 
into  the  blood  stream,  produces  after  three  or  more  weeks  artificial 
tuberculosis  in  guinea-pigs  and  rabbits.  Dogs  and  cats  can  also  be 
infected  by  experimental  inoculation. 

When  a  trace  of  tubercular  virus  is  inserted  subcutaneously  in 
the  thigh  of  a  guinea-pig,  in  about  a  week  or  ten  days  a  chain  of 
enlarged  glands  will  be  easily  felt  in  the  vicinity  of  the  seat  of 
inoculation.  This  affords  an  unfailing  test,  which  can  be  applied 
when  there  is  difficulty  in  ascertaining  by  the  microscope  the  presence 
of  the  bacilli  in  the  material  under  examination.  It  also  affords  a 
valuable  method  for  testing  the  effects  of  antiseptics  on  tubercular 
virus.  The  appearances  observed  at  the  autopsy  are  swollen 
lymphatic  glands,  in  the  neighbourhood  of  the  inoculation,  followed 
by  softening  and  abscess  ;  enlargement  of  the  spleen  and  liver,  with 
formation  of  caseous  tubercles  ;  and  tubercular  deposits  in  the  lungs, 
bronchial  glands,  and  peritoneum. 

After  inoculation  of  the  eye,  grey  tubercles  appear  on  the  iris, 
and  undergo  enlargement  and  caseation,  followed  by  tuberculosis 
of  the  eyeball  and  organs  generally. 

The  bacilli  appear  to  be  the  direct  cause  of  tuberculosis,  and 
the  presence  of  the  bacillus  in  the  sputum  of  patients  is  a  distinctive 
sign  of  the  existence  of  this  disease.  The  detection  of  the  bacillus 
has,  consequently,  become  a  test  which  is  constantly  applied. 

The  bacilli  are  found  in  all  tubercular  growths  of  man,  monkeys, 
cattle  (Perlsuckt),  birds,  and  many  other  animals,  and  in  cases  of 
artificial  tuberculosis,  in  rabbits,  guinea-pigs,  cats,  etc.  In  man  the 
bacillus  can  be  detected  in  the  tissues,  in  the  sputum,  in  the  blood, 
and  in  the  urine. 

Tuberculosis  may  also  be  produced  by  inhalation  and  feeding 
experiments.  The  channels  of  infection  in  man  are  also  most 
probably  the  pulmonary  and  intestinal  mucous  membranes.  The 
possibility  of  inoculation  of  skin  wounds  is  open  to  doubt.  The 
bacilli  or  their  spores  are  inhaled  from  the  air,  or  taken  in  with 
food.  Morphologically  identical  bacilli  have  also  been  observed,  but 
very  sparsely,  in  sections  of  lupus. 

METHODS  OF  EXAMINING  THE  TUBERCLE  BACILLUS. 

Numerous  methods  have  been  recommended  for  examining  the 
tubercle  bacillus.  A  few  of  these  will  be  described,  as  many  are 
only  of  historical  interest. 


TUBERCULOSIS.  385 

The  Ziehl-Neelsen  method  is  preferred  by  the  author  both  for 
sections  and  cover-glass  preparations. 


KocK*  ni'iijhinl  method.  —  Cover-glass  preparations  or  sections  are  laid 
in  Koch's  solution  (No.  23,  r)  for  twenty-four  hours,  or  for  one  hour  if 
the  solution  is  warmed  to  40°  C.  Rinse  in  water  ;  immerse  in  a  watery 
solution  of  vesuvin  for  two  minutes  ;  rinse  again  in  water,  and  examine  ; 
or,  after  rinsing  in  water,  treat  with  alcohol,  clove-oil,  and  Canada 
balsam. 

rJn-HcTC*  method.  —  Cover-glass  preparations  are  allowed  to  float  in  a 
watch-glass,  containing  a  solution  of  gentian-violet  or  fuchsine,  added  to 
aniline  water.  A  saturated  alcoholic  solution  of  the  dye  is  added  till 
precipitation  commences  (10  cc.  aniline  water,  and  10  to  20  drops  of  the 
colour  solution).  The  cover-glasses  are  left  in  the  solution  for  about 
half  an  hour  ;  then  washed  for  a  few  seconds  in  strong  nitric  acid  (one 
part  commercial  nitric  acid  to  two  of  distilled  water),  and  rinsed  in 
distilled  water.  After-stain  with  vesuvin  or  methylene-blue,  rinse  in 
water,  dry  and  preserve  in  Canada  balsam. 
.  Ehrlich-Koch  method. 

Saturated  alcoholic  solution  of  methyl-  violet  or  fuchsine    11 
Aniline  water         ........     100 

Absolute  alcohol    ........       10 

Preparations  are  left  for  twelve  hours  in  this  solution  (colouring  of 
the  cover-glass  preparations  can  be  expedited  by  warming  the  solution). 

Treat  the  preparations  with  (1  to  3)  solution  of  nitric  acid  a  few 
seconds. 

Wash  in  alcohol  (GO  per  cent.)  for  a  few  minutes  (cover-glass  prepara- 
tions need  only  be  rinsed  a  few  times).  After-stain  with  diluted  solution 
of  vesuvin  or  methylene-blue  for  a  few  minutes. 

Wash  again  in  60  per  cent,  alcohol,  dehydrate  in  absolute  alcohol. 
Clear  with  cedar-oil,  mount  in  Canada  balsam. 

Rindfteisch's  method.  —  Prepare  a  solution  composed  of 

Saturated  alcoholic  solution  of  fuchsine     .         .         10  drops 
Aniline  water     .......  2  drams. 

Pour  it  into  a  watch-glass,  and  float  the  cover-glass  ;  warm  the  watch- 
glass  over  a  spirit-lamp  until  steam  rises.  Remove  it  from  the  flame, 
and  set  it  aside  for  five  minutes.  Take  out  the  cover-glass,  and  transfer 
it  for  a  few  seconds  to  acidulated  alcohol  (two  drops  of  nitric  acid  in  a 
watch-glass  full  of  alcohol).  Wash  in  distilled  water,  dry,  and  preserve 
in  balsam.  After-stain,  if  necessary,  with  Bismarck-brown,  or  methylene- 
blue. 

Gibbet  n«t  //<><}.—  Cover-glass  preparations  are  placed  in  Gibbes'  double- 
staining  solution  which  has  been  warmed  in  a  test-tube,  and,  as  soon  as 
steam  rises,  poured  into  a  watch-glass.  They  are  allowed  to  remain  for 
five  minutes,  and  then  are  washed  in  methylated  spirit  till  no  more  colour 
comes  away,  dried  in  the  air  or  over  a  spirit-lamp,  and  mounted  in 
Canada  balsam,  Jf  the  solution  is  used  without  warming,  the  cover-glasses 


386  INFECTIVE   DISEASES. 

must  be  left  in  it  for  an  hour.  Sections  are  treated  on  the  same 
principles,  but  must  be  left  in  the  solution  for  several  hours.  The 
crumpling  of  the  sections  by  the  action  of  nitric  acid  is  avoided. 

Bawngdrten's  method. — Cover-glass  preparations  of  sputum  are  made 
as  already  described,  and  immersed  in  a  very  dilute  solution  of  potash 
(1  to  2  drops  of  a  33  per  cent,  solution  of  potash  in  a  watch-glass  of  dis- 
tilled water).  The  cover-glass  is  pressed  down  on  a  slide,  and  examined 
with  a  high  power.  The  bacilli  can  be  thus  examined  in  the  unstained 
condition,  and  to  avoid  any  mistake  from  confusion  with  other  species, 
the  cover-glass  can  be  removed,  dried,  passed  through  the  flame,  and 
stained  with  a  drop  of  an  aqueous  solution  of  fuchsine,  or  gentian-violet. 
The  putrefactive  bacteria  are  stained,  but  the  tubercle  bacilli  remain 
absolutely  colourless. 

Baumgarten's  new  method. — A  solution  is  prepared  as  follows  :  Drop 
4  to  5  drops  of  concentrated  alcoholic  methyl-violet  solution  into  a  small 
watch-glass  full  of  water,  (a)  Stain  the  sections  in  this  solution,  wash 
them  in  water,  and  decolorise  in  absolute  alcohol  (five  to  ten  minutes)  ; 
or,  before  treating  with  alcohol,  immerse  the  sections  for  five  minutes  in 
a  half- saturated  solution  of  carbonate  of  potash.  Pass  through  clove-oil, 
and  mount  in  a  mixture  of  Canada  balsam,  free  from  chloroform,  and 
clove-oil  (equal  parts).  The  object  of  this  process  is  to  differentiate  the 
tubercle  bacilli  from  chance  bacteria,  inasmuch  as  the  tubercle  bacilli 
are  gradually  decolorised  by  the  clove-oil.  (6)  Sections  stained  in  the 
above  solution  are  placed  for  five  minutes  in  alcohol,  and  then  in  a 
concentrated  solution  of  Bismarck-brown  in  1  per  cent,  solution  of  acetic 
acid.  The  after-treatment  may  be  conducted  as  already  described. 

Ziehl-Neelsen  method. — Cover-glass  preparations  may  be  quickly  stained 
in  Neelsen's  solution  warmed  in  a  watch-glass  till  steam  rises.  Sections 
are  left  for  from  five  to  ten  minutes  in  the  solution,  and  then  washed  in  a 
watery  solution  of  sulphuric  acid  (25  per  cent.),  rinsed  in  distilled  water, 
and  immersed  in  methylene-blue  solution.  After  two  or  three  minutes 
they  are  passed  through  alcohol  and  oil  of  cloves,  and  mounted  in  Canada 
balsam. 

FranlceVs  method. — Sputum  preparations  are  rapidly  double-stained 
by  the  following  method  :  Prepare  a  solution  by  adding  concentrated 
alcoholic  methyl-violet  or  fuchsine  solution,  drop  by  drop,  till  opalescence 
arises,  to  5  ccm.  of  aniline- water  heated  to  100°  C.  Float  the  prepared 
cover-glasses  two  minutes  in  the  warmed  solution.  The  process  of  after- 
staining  and  decolorisation  is  effected  by  placing  the  preparation  for  one 
to  two  minutes  in  one  of  the  following  solutions  :  for  fuchsine-stained 
preparations,  a  saturated  solution  of  methylene-blue  in  a  mixture  of 

Alcohol 50 

Distilled  water .30 

Nitric  acid 20 

which  is  filtered  before  use  ;  for  preparations  stained  in  methyl- violet,  a 
saturated  solution  of  vesuvin  may  be  used  in 

Alcohol .70 

Nitric  acid   ...  30 


TUBERCULOSIS. 


387 


Ehrlich's  Method  and  Eosin. — The  author  has  found  that  after  sections 
have  been  stained  with  methyl-violet  and  Bismarck-brown  by  Ehrlich's 
method,  as  described  by  Koch,  they  may  with  advantage  be  immersed  in 
a  weak  alcoholic  solution  of  eosin,  then  rinsed  in  clean  absolute  alcohol, 
clarified  with  clove-oil,  and  mounted  in  Canada  balsam.  The  giant  cells 
are  then  stained  pink,  while  their  nuclei  are  brown,  and  the  bacilli  blue. 

TUBERCULOSIS  IN  MAN. 

The  disease  manifests  itself  in  various  forms  in  man,  and  most 
frequently  in  the  lungs,  producing  phthisis  or  consumption.  The 
sputum  contains  the  bacilli  in  large  numbers,  and  is  extremely 
virulent.  Scrofula  and  lupus  are  forms  of  tuberculosis ;  they  are 


FIG.  168.— SECTION  THKOUGH  A  LUPUS  NODULE  OF  THE  NOSK. 

probably  produced  by  an  attenuated  variety  of  the  tubercle  bacillus. 
Lupus  can  be  distinguished  from  tuberculosis  of  the  skin ;  and 
scrofulous  lymphatic  glands  are  distinguished  from  tubercular  glands 
by  the  tendency  of  the  latter  to  produce  generalised  tuberculosis. 
This  difference  in  the  intensity  of  the  virus  in  the  two  cases, 
Lingard  illustrated  by  the  effect  upon  inoculated  guinea-pigs. 

Cavities  in  the  lungs  are  often  thickly  lined  with  bacilli.  They 
are  present  in  great  numbers  in  the  caseous  matter,  though  in 
equine  and  bovine  tuberculosis  this  is  not  the  case. 

Whether  the  disease  in  man  is  contagious  is  an  open  question, 
though  numerous  cases  of  supposed  communication  between  husband 
and  wife,  brothers  and  sisters,  have  been  reported,  and  Ransome 


388 


INFECTIVE   DISEASES. 


showed  that  tubercle  bacilli  were  present  in  the  breath  in  phthisis. 
On  the  other  hand,  the  experience  in  consumption  hospitals  does  not 


FIG.  169.— TUBERCULAR  ULCERATION  IOP  MUCOSA  OF  HUMAN  ILEUM. 
Between  the  ulcers  there  are  tubercular  lymph-follicles  (HAMILTON). 

support  this  view,  there  being  no  evidence  of  the  communication  of 
the  disease  to  nurses  and  hospital  attendants, 


TUBERCULOSIS. 


389 


TUBERCULOSIS  OF  CATTLE. 

In  cattle  the  disease  may  occur  as  the  result  of  inhaling  bacilli, 
or  of  ingestion  with  food.  It  is  very  frequently  found  in  the  lungs ; 
and  calves  may  be  infected  by  milk  from  cows  with  tubercular 
udders.  Calves  may  also  suffer  from  congenital  tuberculosis,  the 
bacilli  having  been  transmitted  from  the  mother  during  gestation. 

Breeding  in-and-in,  over-production  of  milk,  and  confinement  with 
insanitary  surroundings,  predispose  to  tuberculosis.     The  disease  is 
known  in  Germany  as  "  Perlsucht " ;  and  in  this  country  the  lesions 
on  the  pleura  are  known  as 
"grapes,"    and    the    animals 
themselves      are      commonly 
called  "  wasters." 

The  disease  may  also  exist 
in  the  lungs  or  in  other 
organs,  in  a  limited  form, 
without  any  indication  of 
ill  health.  In  such  cases 
the  disease  can  be  detected 
by  injection  of  tuberculin,  a 
marked  rise  of  temperature 
occurring  in  t  u  b  e  r  c  u  1  a  r 
animals. 

In  advanced  cases,  the 
symptoms  commonly  observed 
are  cough,  difficulty  in  breath- 
ing, staring  coat,  wasting,  and 
diarrhoea  ;  and  if  the  udder  is  infected,  nodules  in  the  gland,  arid  thin 
bluish  milk.  In  the  lungs,  after  slaughter,  a  few  small  cheesy  tubercles 
may  be  found  in  animals  apparently  in  perfect  health  and  in  prime 
condition  for  the  market.  In  advanced  cases,  the  lungs  on  section 
show  large  yellow  masses,  containing  calcified  matter,  and  the 
bronchi  may  be  full  of  yellowish  pasty  contents.  The  disease  will  be 
found  to  involve  the  bronchial  glands.  The  serous  membrane  may 
be  covered  with  little  warts  or  grape-like  masses.  The  lymphatic 
glands  may  be  enlarged  to  an  enormous  size.  Tubercular  ulceration 
of  the  intestine  is  sometimes  found,  but  not  commonly.  In  tubercular 
disease  in  the  udder,  a  painless  swelling  is  found  which  may  affect 
one  or  more  quarters  of  the  gland. 

Transmission  of  Tuberculosis  from  Man  to  Cattle. — It  is 


FIG.  170. —SECTION  OF  LUPUS  OF  THE  SKIN, 
x  700.  Giant  cell  containing  a  tubercle 
bacillus  (FLUGGE). 


390 


INFECTIVE    DISEASES. 


for  obvious  reasons  impossible  to  ascertain  by  experiment  whether 
tuberculosis  can  be  transmitted  from  cows  to  man  by  milk  or  other- 
wise ;  but  some  light  may  be  thrown  upon  this  important  question 
by  ascertaining  the  result  of  inoculating  bovines  with  human  tuber- 
culosis. If  calves  can  be  infected  with  tuberculosis  from  a  human 
source  by  inoculation  or  ingestion  experiments,  and  especially  if 
the  effect  of  administering  human  and  bovine  tubercle  to  calves 
by  these  means  is  found  to  be  the  same,  such  experiments  will 
not  only  serve  to  dispel  any  doubt  there  may  be  as  to  the  identity 
of  the  two  affections,  but  they  will  strengthen  the  hands  of  those 


FIG.  171.— TUBERCULOSIS  OF  PLEURA  ;  ".GRAPE- DISEASE." 

who  insist  upon  the  necessity  of  more  thorough  inspection  of  dairy 
cows,  and  of  power  to  deal  with  tubercular  animals. 

Inoculation  of  a  Calf  with  Human  Tubercular  Sputum. — The 
author  obtained  sputum  containing  numerous  bacilli  from  an 
advanced  case  of  phthisis.  The  sputum  was  shaken  up  with  sterilised 
salt  solution  and  injected  into  the  peritoneal  cavity.  A  few  weeks 
afterwards  the^calf  showed  signs  of  illness.  The  animal  looked  dull, 
did  not  feed  well,  had  a  slight  cough,  and  showed  less  inclination  to 
move  about  than  usual.  These  symptoms  gradually  increased,  and 
death  occurred  forty-two  days  after  inoculation.  Extensive  lesions 


TUBERCULOSIS.  391 

were  discovered  at  the  post-mortem  examination.  The  mesentery 
was  adherent  to  the  abdominal  wall,  at  the  seat  of  the  inoculation, 
and  to  the  rumen  ;  the  liver  was  adherent  to  the  diaphragm.  There 
was  extensive  tubercular  deposit  at  the  seat  of  inoculation,  and 
an  abscess  the  size  of  a  walnut.  Extending  over  the  mesentery 
from  this  point  there  were  hundreds  of  wartlike,  fleshy,  new  growths, 
some  quite  irregular  in  form,  others  spherical  or  button-shaped. 
There  were  similar  deposits  on  the  under  surface  of  the  liver,  on  the 
spleen,  in  the  gastro-splenic  omentum,  and  on  the  peritoneal  surface 
of  the  diaphragm.  The  spleen  was  adherent  to  the  rumen,  and 
on  dissecting  away  the  adhesions  another  abscess  was  opened.  The 
lungs  were  congested  and  the  pleurae  thickened.  On  microscopical 
examination  of  sections  extremely  minute  tubercles  were  found  to 
be  disseminated  throughout  the  whole  of  the  substance  of  the  lungs 
and  liver,  and  tubercle  bacilli  were  found  in  these  and  in  the 
peritoneal  deposits.  The  abscesses  contained  Streptococcus  pyogenes. 
The  calf  died  of  pyaemia,  but  sufficient  time  had  elapsed  for  marked 
local  infection  leading  to  generalised  miliary  tuberculosis. 

TUBERCULOSIS  IN  RELATION  TO  THE  PUBLIC  MILK  SUPPLY. 

There  is  not  the  slightest  doubt  that  when  the  udder  is  involved 
the  milk  is  highly  virulent  to  the  lower  animals,  and  presumably 
is,  therefore,  dangerous  to  man.  The  virulence  of  the  milk  was 
first  insisted  upon  by  Klencke  in  1846,  and  confirmed  by  Gerlach  in 
1869,  and  later,  by  others. 

This  subject  was  again  brought  forward  with  the  discovery  of 
the  tubercle  bacillus,  and  the  demonstration  of  its  existence  in  the 
milk  in  certain  cases  of  bovine  tuberculosis.  Koch  pointed  out  that 
the  milk  only  contained  bacilli,  and  was  only  infective,  when  the 
udder  itself  was  tubercular.  By  this  he  explained  the  contradictory 
results  obtained  by  various  experimenters  with  milk  from  cows  un- 
doubtedly suffering  from  "  Perlsucht."  Koch  considered  that  positive 
effects  were  obtained  with  milk  when  it  happened  to  contain  tubercle 
bacilli,  and  negative  with  milk  from  which  they  were  absent.  Bang 
in  a  number  of  cases  verified  the  presence  of  tubercle  bacilli  in  milk, 
and,  owing  to  the  contradictory  results  of  previous  investigations, 
repeated  the  ingestion  experiments.  The  milk  was  found  to  be 
virulent  both  to  pigs  and  rabbits. 

In  this  country  Woodhead  and  M'Fadyean  tested  milk  for 
tubercle  bacilli.  They  examined  six  hundred  cows  in  the  Edinburgh 
dairies,  and  found  thirty-seven  suffering  from  mammitis,  but  in  only 


392  INFECTIVE    DISEASES. 

six  were  they  able  to  demonstrate  the  presence  of  tubercle  bacilli  in 
the  milk,  and  then  only  in  small  numbers. 

Hirschberger  found  in  twenty  cases  of  tuberculosis  in  cattle 
that  the  milk  of  eleven  was  virulent  to  guinea-pigs.  Three  cows  out 
of  nine  in  which  the  disease  was  restricted  to  the  lungs  gave  infected 
milk.  On  the  other  hand,  Nocard  inoculated  milk  from  eleven 
tuberculous  cows,  of  which  only  one  had  diseased  udder,  and  only 
this  one  gave  infective  milk.  Bang  injected  rabbits  with  milk  from 
twenty-one  cases  of  tuberculosis,  with  the  udders  apparently  normal, 
and  the  milk  was  virulent  in  two. 

The  author  had  two  cases  of  udder  tuberculosis  under  observation, 
and  as  no  experiments  had  at  the  time  been  made  in  this  country 
with  milk  known  to  contain  tubercle  bacilli,  it  was  decided  to  study 
the  effect  on  rabbits,  and  test  the  results  obtained  by  Bang.  These 
cases  were  both  interesting  and  instructive,  and  may  be  referred  to 
in  detail. 

One  was  a  case  of  advanced  general  tuberculosis.  There  was  extreme 
emaciation,  general  apathy,  and  a  peculiar  dull  expression  of  countenance. 
The  skin  was  dry  and  harsh,  the  coat  staring,  and  there  was  loss  of  hair 
in  patches  about  the  face  and  neck.  There  was  dulness  on  percussion 
over  a  large  area  of  the  thorax,  and  the  respirations  were  increased  in 
rapidity.  There  was  also  occasional  cough  and  some  diarrhoea.  But  the 
most  interesting  condition  was  observed  on  examination  of  the  udder. 
The  gland  was  swollen,  especially  posteriorly,  and  distinct  induration 
could  be  felt  on  examination.  The  deposit  appeared  to  be  more  or  less 
limited  to  the  posterior  quarters.  The  cow  evinced  no  pain  during  the 
examination  of  the  udder,  not  even  on  the  application  of  firm  pressure. 

The  author  took  samples  in  test-tubes  of  the  milk  from  all  four  teats  ; 
when  freshly  drawn,  it  differed  noticeably  from  the  normal  secretion.  It 
was  a  thin,  watery,  turbid  fluid  with  whitish  flakes  in  suspension,  but  it 
was  not  gelatinous  or  muco-purulent  in  character,  and  was  free  from  any 
markedly  yellow  colour.  After  being  set  aside  in  the  laboratory  for 
some  hours  it  separated  into  a  layer  of  cream  and  a  turbid  liquid  of  a 
yellowish  tint,  while  at  the  bottom  of  the  test-tube  there  was  a  whitish 
flocculent  deposit,  especially  in  the  samples  from  the  posterior  quarters. 

There  were  tubercle  bacilli  both  in  the  cream  and  in  the  deposit.  In 
the  cream  they  were  only  present  in  small  numbers,  and  were  detected, 
therefore,  only  after  careful  search.  But  in  the  deposit  they  were  readily 
found,  as  in  a  cover-glass  preparation  there  were  sometimes  four  or  five 
in  the  field  of  the  microscope. 

The  method  adopted  for  the  examination  of  this  deposit  was  as 
follows  :  The  whole  of  the  liquid  in  the  test-tube  was  carefully  poured 
off,  and  a  trace  of  the  sediment  spread  out  on  a  cover-glass.  This  was 
allowed  to  dry,  and  passed  through  the  flame,  and  stained  in  hot  Ziehl- 
Neelsen  solution  in  the  usual  manner. 


TUBERCULOSIS. 


393 


The  other  cow  was  also  a  case  of  general  tuberculosis,  and  presented 
somewhat  similar  lesions  of  the  udder.  The  induration  of  the  gland  was 
readily  detected,  and  examination  of  the  milk  showed,  as  in  the  previous 
case,  the  presence  of  tubercle  bacilli. 

It  will  be  observed  that  in  neither  of  these  cases  was  the  disease 
limited  to  the  udder  ;  in  both  the  implication  of  the  gland  was  part  of 
general  tuberculosis. 


FIG.  172.— TUBERCULAR  ULCERATION  OF  THE  INTESTINE  OF  A  Cow. 

The  first  cow  was  killed,  and  the  following  lesions  were  found  at  the 
post-mortem  examination. 

T/nn'dx. — The  lungs  and  bronchial  glands  were  extensively  invaded 
with  tubercular  deposit.  The  glands  were  greatly  enlarged  and  densely 
fibrous,  in  many  cases  with  central,  stone-like  masses,  grating  on  section 
against  the  edge  of  the  knife.  In  the  lung  there  was  every  stage,  from 
the  early  deposit  to  purulent  cavities,  cheesy  masses,  and  calcified  dtbrt*. 

Abdomen. — There  were  a  few  caseous  nodules  in  the  liver,  but  none 
in  the  spleen.  The  mesenteric  glands  formed  an  almost  continuous  chain 


394  INFECTIVE    DISEASES. 

of  large  tumours,  mostly  with  central  cretification.  Tubercular  deposit 
in  the  intestines  could  be  recognised  from  the  outside,  and  on  laying 
them  open  the  mucous  membrane  was  found  to  be  studded  with  tuber- 
cular ulcers.  These  ulcers  were  most  numerous  in  the  large  intestine, 
and  varied  in  size  from  a  sixpence  to  a  florin.  Some  were  circular,  others 
slightly  irregular  in  form,  and  others  again  distinctly  oval.  In  the  latter 
case  they  were  generally  situated  with  their  long  diameter  transversely. 
The  base  of  the  ulcer  involved  the  muscular  coat,  and  was  irregularly 
radiated.  The  margin  was  broad,  and  elevated  above  the  general  surface, 
producing  a  ring-like  appearance. 

Mammary  Gland. — The  udder  was  infiltrated  throughout  with  tuber- 
cular new  growth,  but  the  invasion  was  most  marked  in  the  posterior 
quarters.  There  was  apparently  very  little  tendency  to  caseation. 

Microscopical  Examination  of  the  Udder. — In  order  to  study  the  histo- 
logical  characters  of  the  gland,  and  the  distribution  of  the  bacilli,  sections 
were  stained  with  logwood  and  rubin,  and  others  again  with  fuchsine 
and  methylene-blue.  The  tubercular  new  growth  consisted  of  the  usual 
histological  elements,  round  cells,  epithelioid  cells,  and  giant  cells. 
Healthy  lobules  here  and  there  were  sharply  marked  off  from  those  in 
which  the  growth  was  compressing  and  obliterating  the  alveoli  in  its 
progress.  Bacilli  were  present  in  the  giant  cells,  and  also  distributed  in 
vast  numbers  throughout  the  tubercular  tissue  generally.  Bacilli  were 
found  in  epithelioid  cells  close  to  the  alveolus,  and  also  between  the  cells 
lining  the  alveoli.  In  parts  also  the  new  growth  had  involved  the  milk 
ducts,  and  therefore  it  was  easy  to  account  for  the  presence  of  the  bacilli 
in  the  milk. 

The  bacilli  were  found  in  considerable  numbers  also  in  sections  of  the 
intestinal  ulcers. 


EXPERIMENTAL  INFECTION  OF  BABBITS. 

Ingestion. — A  rabbit  received  the  contents  of  a  test-tube  which 
had  been  filled  with  milk  from  one  of  the  posterior  teats,  mixed  with 
a  small  quantity  of  bran.  In  four  weeks  there  was  commencing 
emaciation ;  later,  diarrhoea  set  in,  and  death  occurred  exactly  fifty- 
eight  days  after  administration  of  the  milk.  At  the  post-mortem 
examination  the  mesenteric  glands  were  found  to  be  much  enlarged 
and  caseous.  A  cover-glass  preparation  from  a  crushed  gland 
revealed  numerous  tubercle  bacilli.  On  opening  the  intestines  there 
was  a  patch  of  ulceration,  showing  the  point  of  access  of  the  bacilli. 
The  intestinal  ulceration  was  a  reproduction,  to  a  certain  extent,  of 
the  condition  in  the  cow  which  had  been  the  source  of  the  virus. 

Subcutaneous  Injection. — A  second  rabbit  was  injected  under  the 
skin  of  the  back  by  means  of  a  capillary  pipette  with  about  ten 
drops  of  milk,  including  some  of  the  deposit  from  the  bottom  of  the 
test-tube.  The  sample  of  milk  had  in  this  case  also  been  taken  from 


DESCRIPTION    OF    PLATE    XII. 
Tubercular  Mammitis. 

FIG.  .1.— From  a  section  of  the  udder  of  a  milch  cow.  The  tubercular  deposit 
is  seen  to  invade  the  lobules  of  the  gland.  Lobules  comparatively  healthy 
are  marked  off,  more  or  less  sharply,  from  the  diseased  ones  in  which  the 
new  growth  in  its  progress  compresses  and  obliterates  the  alveoli.  Stained 
by  the  Ziehl-Neelsen  method  and  with  methylene-blue.  x  50. 

FIG.  2.— Part  of  the  same  preparation.  On  the  right  of  the  section  part  of  a 
healthy  lobule  is  seen.  On  the  left  a  lobule  is  invaded  by  tubercular  new 
growth  composed  of  round  cells,  epithelioid  cells  and  typical  giant  cells. 
Tubercle  bacilli  can  be  seen  both  singly  and  collected  in  groups.  They 
are  found  in  and  between  the  cells,  and  in  the  interior  of  giant  cells. 
Bacilli  may  be  seen  between  the  cells  lining  an  alveolus  and  projecting 
into  its  lumen,  x  800. 


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Ftfl. 


Fig  2. 

TUBERCULAR   MAMMITIS 


E.Jf.CrooJwluinJe/ecit 


TUBERCULOSIS. 


395 


one  of  the  posterior  teats.  The  rabbit  was  placed  in  a  separate 
hutch,  and  death  from  general  tuberculosis  occurred  ninety- two  days 
after  inoculation. 

The  diaphragm  and  mesentery  were  studded  with  tubercles  the 
size  of  a  pin's  head.  The  kidneys  superficially  showed  whitish 
rounded  nodules  projecting  above  the  surface.  These  were  found 
on  section  to  be  continuous,  with  wedge-shaped  deposits  in  the  sub- 
stance of  the  kidney.  The  lungs  presented  a  very  striking  appear- 
ance, being,  in  short,  a  mass  of  tubercular  deposit ;  and  the  bronchial 
and  tracheal  glands  were  similarly  affected.  In  sections  of  the 
kidney  and  lung  the  bacilli  were  present,  but  they  were  distributed 
irregularly ;  in  one  part  of  a  section  it  was  difficult  to  detect  a 
single  bacillus,  in  other  parts  they  were  present  in  large  numbers. 

The  milk  from  the  two  cows, 
previously  to  their  coming  under 
observation,  had  been  mixed  with 
the  general  supply  of  a  dairy.  There 
is  indeed  ample  evidence  that,  both 
in  this  and  in  other  countries,  the 
milk  of  tuberculous  animals  finds  its 
way  into  the  market.  The  question 
which  naturally  arises  is  the  possi- 
bility of  any  manifestation  of  tuber- 
culosis in  man,  arising  from  the 
consumption  of  unboiled  milk  con- 
taining tubercle  bacilli.  We  must 
admit  that  there  is  no  direct 
evidence  of  the  transmission  of 
tuberculosis  by  milk  from  cow  to 
man ;  but  this  may  arise  from  the  difficulty  in  tracing  such  a 
source  of  infection,  owing  to  the  long  time  which  elapses  before 
symptoms  manifest  themselves  in  man.  Yet,  if  milk  be  a  source  of 
infection,  we  should  naturally  expect  that  primary  tuberculosis  of  the 
intestine  would  be  by  no  means  an  uncommon  manifestation  of  the 
disease;  and  this  in  the  adult  is  not  in  accordance  with  clinical 
experience.  Such  an  argument  would  tend  to  centra-indicate 
danger  to  adults;  but,  on  the  other  hand,  the  possible  danger  to 
children  has  been  rightly  insisted  upon  by  the  earliest  writers  on 
this  subject.  Woodhead  has  recently  stated  that,  from  his  experi- 
ence in  two  large  hospitals,  he  has  been  much  struck  by  the  fact 
that,  in  children  who  had  died  from  other  diseases  during  the  course 
of  tubercular  disease  of  the  abdominal  glands,  there  was  frequently 


FIG.  173.— TUBERCULAR  ULCERA- 
TION  OF  THE  INTESTINE  OF  A 
RABBIT. 


396 


INFECTIVE    DISEASES. 


not  any  trace  of  tubercular  disease  in  other  parts ;  thus  pointing  to 
the  intestine  as  the  channel  by  which  the  bacillus  made  its  way  into 
the  body.  Woodhead  also  remarks  that  in  a  large  number  of  cases 


FIG.  174.— TUBERCULOSIS  OF  THE  LUNGS. 

From  a  photograph  of  the  lungs  of  a  rabbit  which  had  been  injected  sub- 
cutaneously  with  about  ten  drops  of  milk,  including  in  suspension  a  small 
quantity  of  the  deposit  at  the  bottom  of  a  sample  of  milk  from  a  cow  with 
tuberculosis  of  the  udder.  Death  occurred  from  general  tuberculosis  ninety-two 
days  afterwards.  The  appearance  of  the  lungs  was  very  striking.  They  were 
almost  completely  composed  of  tubercular  deposit.  The  bronchial  glands, 
as  well  as  the  tracheal,  of  which  one  is  seen  in  the  photograph,  were  also 
enlarged  and  caseous.  There  were  tubercular  deposits  in  the  kidneys  and 
other  organs,  and  also  at  the  seat  of  inoculation. 

of  general  tuberculosis,   where   the  possibility  of  infection  by  the 
pulmonary  passages  was  evidently  excluded,  the  tubercular  process 


TUBERCULOSIS.  397 

appeared  to  have  invaded  the  body  by  the  intestinal  canal.  These 
facts,  taken  in  connection  with  the  occasional  existence  of  tubercle 
bacilli  in  milk,  went  far  to  prove,  in  his  opinion,  that  milk  was  a 
source  of  tubercular  infection,  especially  to  young  children. 

From  his  own  experiments  and  observations  the  author  has 
drawn  the  following  conclusions  : — 

1.  Cows  with  tuberculosis  of  the  udder  are  to  be  found  in  dairies 

in  this  country. 

2.  The  milk  of  these  cows  is,  as  a  rule,  mixed  with  the  general 

supply. 

3.  The   milk   in   cases  of   udder   tuberculosis   contains   tubercle 

bacilli. 

4.  Rabbits  inoculated  with,  or  fed  upon,  milk  containing  tubercle 

bacilli  contract  tuberculosis. 

5.  Direct  evidence  of  transmission  of  tuberculosis  by  milk  to  man  is 

wanting,  but  from  the  effect  of  such  milk  on  the  lower  animals 
it  is  reasonable  to  conclude,  in  the  present  state  of  our  know- 
ledge, that  there  may  be  danger  in  using  the  milk  of  cows 
with  tubercular  udders,  and  therefore  strict  inspection  of 
dairies  should  be  enforced  ;  and  boiling  of  milk  before 
use  will,  as  a  rule,  be  a  wise,  if  not  absolutely  a  necessary 
precaution. 

Bellinger  has  shown  that  the  virulence  of  cow's  milk  is  reduced 
by  dilution  with  water  in  the  proportion  of  1  in  40  and  even  of 
1  in  100,  and  that  therefore  there  would  be  much  less  danger 
in  consuming  tubercular  milk  which  had  been  mixed  with  the 
milk  of  healthy  cows,  than  there  would  be  in  taking  it  direct 
from  the  infected  cow.  This  is  a  matter  of  scientific  interest ;  but 
it  would  be  no  justification  for  a  dairyman  to  mix  the  milk  of  a 
tubercular  cow  with  milk  of  cows  known  to  be  healthy.  The  milk 
of  cows  suffering  from  tuberculosis  should  undoubtedly  be  rejected. 


TUBERCULOSIS  AND  THE  PUBLIC  MEAT  SUPPLY. 

The  question  of  the  advisability  of  allowing  the  flesh  of  tuber- 
cular animals  to  be  sold  for  food,  especially  when  the  disease  exists 
in  a  very  small  degree,  is  a  vexed  one.  Numerous  experiments 
have  been  made  upon  the  infectivity  of  the  flesh  of  tubercular 
animals.  Kastner  inoculated  the  juice  expressed  from  the  flesh  of 
tubercular  cows.  Sixteen  guinea-pigs  were  unaffected  after  injection 


398  INFECTIVE   DISEASES. 

of  1  to  2  cc.  into  the  peritoneal  cavity.  Nocard  injected  ten  to 
twenty  drops  of  the  muscle  juice  of  the  hearts  of  tubercular  cattle,  in 
which  the  disease  was  well  marked,  and  none  of  the  guinea-pigs  were 
infected.  With  juice  of  the  muscles  of  the  thigh  derived  from  ten 
tubercular  cows  Nocard  inoculated  forty  guinea-pigs,  and  one  only 
showed  signs  of  tubercle.  Nocard  concluded  that  if  there  was  any 
danger  in  the  flesh  of  tuberculous  animals,  it  was  the  exception  and 
not  the  rule.  On  the  other  hand,  Chauveau  and  Arloing  produced 
tuberculosis  in  two  guinea-pigs  out  of  ten  inoculated  with  muscle 
juice  from  a  tubercular  steer. 

In  1890  a  Royal  Commission  was  appointed  to  investigate  this 
subject,  and  the  report  was  issued  in  1895.  Martin,  on  behalf  of 
the  Commission,  tested  the  flesh  of  twenty-one  tubercular  cows.  In 
two  cases  only  was  evidence  obtained  of  the  presence  of  the  bacillus 
by  inoculation  of  guinea-pigs.  The  flesh  of  eight  cows  affected  with 
mild  tuberculosis  produced  tubercle  in  one  instance  by  inoculation, 
but  the  ingestion  experiments  were  negative.  The  flesh  of  five  cows 
severely  affected  with  tubercle  gave  the  disease  in  four  cases,  either 
by  feeding  or  inoculation,  but  only  one  gave  the  disease  both  ways. 
Martin  thought  that  some  of  the  results  were  due  to  the  butcher 
infecting  the  meat  in  the  process  of  dressing  the  carcase,  either  by 
his  hands  or  knives.  Woodhead  made  a  series  of  experiments  to  test 
the  effects  of  roasting  and  boiling  on  the  tubercular  virus  in  meat. 
It  was  found  that  in  boiling  and  roasting  experiments,  as  ordinarily 
carried  out  in  the  kitchen,  the  temperature,  however  high  it  may  be 
on  the  surface,  seldom  reaches  60°  C.  in  the  centre,  except  in  the 
case  of  joints  less  than  about  six  pounds  in  weight.  Boiling  and 
roasting  were  found  insufficient  to  destroy  tubercular  virus  enveloped 
in  rolls  of  meat. 

The  following  were  among  the  conclusions  of  the  Commissioners  :  — 

We  have  obtained  ample  evidence  that  food  derived  from  tuberculous 
animals  can  produce  tuberculosis  in  healthy  animals.  The  proportion  of 
animals  contracting  tuberculosis  after  experimental  use  of  such  food  is 
different  in  one  and  another  class  of  animals ;  both  carnivora  and 
herbivora  are  susceptible,  and  the  proportion  is  high  in  pigs.  In  the 
absence  of  direct  experiments  on  human  subjects,  we  infer  that  man  also 
can  acquire  tuberculosis,  by  feeding  upon  materials  derived  from  tuber- 
culous food-animals. 

The  actual  amount  of  tuberculous  disease  among  certain  classes  of 
food-animals  is  so  large  as  to  afford  to  man  frequent  occasions  for 
contracting  tuberculous  disease  through  his  food.  As  to  the  proportion 
of  tuberculosis  acquired  by  man,  through  his  food  or  through  other  means, 
we  can  form  no  definite  opinion,  but  we  think  it  probable  that  an 


TUBERCULOSIS.  399 

appreciable  part  of  the  tuberculosis  that  affects  man  is  obtained  through 
his  food. 

The  circumstances  and  conditions  with  regard  to  the  tuberculosis  in 
the  food-animal  which  lead  to  the  production  of  tuberculosis  in  man  are, 
ultimately,  the  presence  of  active  tuberculous  matter  in  the  food  taken 
from  the  animal,  and  consumed  by  the  man  in  a  raw  or  insufficiently 
cooked  state. 

Tuberculous  disease  is  observed  most  frequently  in  cattle  and  in 
swine.  It  is  found  far  more  frequently  in  cattle  (full  grown)  than  in 
calves ;  and  with  much  greater  frequency  in  cows  kept  in  town  cow- 
houses than  in  cattle  bred  for  the  express  purpose  of  slaughter.  Tuber- 
culous matter  is  but  seldom  found  in  the  meat  substance  of  the  carcase  ;  it 
is  principally  found  in  the  organs,  membranes,  and  glands.  There  is 
reason  to  believe  that  tuberculous  matter,  when  present  in  meat  sold  to 
the  public,  is  more  commonly  due  to  the  contamination  of  the  surface 
of  the  meat  with  material  derived  from  other  diseased  parts,  than  to 
disease  of  the  meat  itself.  The  same  matter  is  found  in  the  milk  of  cows 
when  the  udder  has  become  invaded  by  tuberculous  disease,  and  seldom 
or  never  when  the  udder  is  not  diseased.  Tuberculous  matter  in  milk  is 
exceptionally  active  in  its  operation  upon  animals  fed  either  with  the 
milk  or  with  the  dairy  produce  derived  from  it.  No  doubt  the  largest 
part  of  the  tuberculosis  which  man  obtains  through  his  food  is  by  means 
of  milk  containing  tuberculous  matter. 

Provided  every  part  that  is  the  seat  of  tuberculous  matter  can  be 
avoided  and  destroyed,  and  provided  care  be  taken  to  save  from  contami- 
nation by  such  matter  the  actual  meat  substance  of  a  tuberculous 
animal,  a  great  deal  of  meat  from  animals  affected  by  tuberculosis  may 
be  eaten  without  risk  to  the  consumer. 

Ordinary  processes  of  cooking  applied  to  meat  which  has  got  con- 
taminated on  its  surface  are  probably  sufficient  to  destroy  the  harmful 
quality.  They  would  not  avail  to  render  wholesome  any  piece  of  meat 
that  contained  tuberculous  matter  in  its  deeper  parts.  In  regard  to  milk 
we  are  aware  of  the  preference  by  English  people  for  drinking  cow's  milk 
raw — a  practice  attended  by  danger,  on  account  of  possible  contamination 
by  pathogenic  organisms.  The  boiling  of  milk,  even  for  a  moment,  would 
probably  be  sufficient  to  remove  the  very  dangerous  quality  of  tuber- 
culous milk. 


TUBERCULOSIS  IN  EQUINES. 

Tuberculosis  is  not  very  common  in  the  horse,  but  when  it  does 
occur,  it  is  frequently  mistaken  for  glanders.  There  may  be  miliary 
tuberculosis  in  the  lungs,  or  nodules  disseminated  throughout  the 
lungs,  liver,  spleen,  and  bones.  In  a  number  of  cases  investigated 
by  Nocard,  the  disease  commenced  in  the  abdominal  organs,  and 
the  affection  of  the  lungs  appeared  to  be  secondary.  The  author 
has  examined  several  cases  of  equine  tuberculosis.  In  some  cases 


400  INFECTIVE    DISEASES. 

the  lungs  were  affected  with  the  disease  in  a  miliary  form.  The 
bacilli  could  not  be  distinguished  from  bacilli  in  sections  of  the  bovine 
disease.  Giant  cells  were  extraordinarily  numerous,  and  in  many 
cases  were  densely  packed  with  bacilli,  so  that  they  could  be  recog- 
nised en  masse  under  a  low  power.  The  bacilli  were  also  distributed 
in  the  tissue  generally,  but  were  much  more  numerous  in  the  giant 
cells. 

TUBERCULOSIS  IN  DOGS. 

Peters  described  a  case  of  tuberculosis  in  a  pet  dog,  from  eating 
sputum  from  a  tubercular  patient.  This  is  said  to  be  a  not 
uncommon  cause  of  canine  tuberculosis. 

TUBERCULOSIS  IN  CATS. 

Nocard  reported  a  case  of  tuberculosis  in  a  cat  from  eating  tuber- 
cular sputum.  The  abdominal  organs  were  diseased.  Bellinger 
has  described  two  cases  of  miliary  tuberculosis.  M'Fadyean  also  has 
described  a  case.  The  bacilli  are  very  plentiful  in  the  lung.  A 
minute  examination  of  the  individual  micro-organisms  by  the  author 
did  not  reveal  any  distinctive  character. 

TUBERCULOSIS  IN  SWINE. 

The  author  examined  the  tubercular  liver  of  a  pig.  The  pig 
was  about  six  months  old,  and  after  suffering  from  cough  and 
emaciation,  died. 

The  liver  had  caseous  nodules  scattered  throughout  its  substance, 
some  the  size  of  a  pea,  and  others  larger.  Tubercle  bacilli  without 
distinctive  characters  were  found  on  examination  of  sections ;  but 
it  was  in  some  parts  of  a  preparation  difficult  to  detect  any  bacilli, 
and  in  other  parts  there  were  not  more  than  five  or  six  in  the 
field  of  the  microscope.  Tuberculosis  in  swine  is  said  to  be  very 
rare  in  America. 

TUBERCULOSIS  IN  BIRDS. 

Hens,  guinea-fowls,  turkeys,  pheasants,  and  partridges,  are  sub- 
ject to  tuberculosis,  and  ostriches  and  other  birds  kept  in  confinement 
may  contract  the  disease. 

Tuberculosis  in  fowls  appears  to  be  introduced  principally  with 
the  food,  the  disease  occurring  commonly  in  the  intestines  and 
liver. 


DESCRIPTION    OF    PLATE    XIII. 
Tuberculosis  in  Swine. 

Section  of  liver  of  a  pig  with  scattered  tubercular  nodules.     Microscopica 
sections  of  the  liver  showed  tubercle  bacilli  in  very  small  numbers. 


Plate  XIII 


C 


Vincent  Broole3.Day  *5o7i,Zit)i. 


TUBERCULOSIS.  401 

The  author  has  examined  several  cases  of  so-called  spontaneous 
tuberculosis  in  fowls.  Sections  of  the  liver  were  in  one  case  remark- 
able on  account  of  the  extraordinary  invasion  of  the  caseous  deposits 
with  bacilli.  Cover-glass  preparations  had  been  made  from  the 
liver  in  the  following  way  for  diagnostic  purposes  :  A  tubercle  was 
readily  picked  out  on  the  point  of  a  scalpel  and  crushed  between  two 
slides,  and  the  cover-glass  preparations  stained  with  the  Ziehl- 
Neelsen  solution.  The  bacilli  are  for  the  most  part  very  small.  A 
few  attain  a  considerable  length,  but  the  majority  are  in  the  form 
of  small,  straight  rods,  with  many  sizes  intervening  between  these 
rods  arid  isolated  granules. 

In  July  1888  the  author  received  from  Mr.  Bland  Sutton  the 
liver  and  lungs  of  a  Rhea,  which  had  died  in  the  Zoological  Gardens. 
The  lung  was  infiltrated  with  caseous  deposits,  and  there  were 
scattered  caseous  nodules  in  the  liver  varying  in  size  from  a  pea 
to  a  marble.  The  naked-eye  appearance  of  a  section  of  the  liver 
through  these  nodules,  at  once  recalled  to  mind  the  naked-eye 
appearance  of  the  deposits  in  the  pig's  liver  already  described.  But 
whereas  in  microscopical  preparations  of  the  pig's  liver,  bacilli  were 
very  scantily  present,  the  sections  of  the  lung  and  liver  of  the  E-hea 
contained  bacilli  in  such  extraordinary  numbers  that,  under  a  power 
of  fifty  diameters,  the  collections  of  bacilli  could  be  recognised  as  red 
granular  masses.  These  red  masses  under  a  high  power  were  re- 
solved into  dense  colonies  of  bacilli.  In  their  number  and  their 
distribution  in  the  tissues,  in  their  varying  size,  and  in  the  extra- 
ordinary length  of  the  longest  forms,  they  presented  very  interesting 
points  for  observation.  From  the  naked-eye  appearance  of  the 
disease  and  the  general  microscopical  characters,  as  well  as  the 
presence  of  bacilli  agreeing  in  their  staining  reactions  with  the 
classical  tubercle  bacilli,  the  author  had  no  hesitation  in  pronouncing 
the  disease  to  be  avian  tuberculosis. 

Klein,  who  had  examined  a  similar  case,  alluded  to  it  in  a 
description  of  leprosy;  but  this  disease  is  unknown  in  the  lower 
animals,  and  all  attempts  to  infect  them  from  man  have  been 
almost,  if  not  entirely,  negative. 

The  bacilli  in  the  Rhea  are  principally  collected  in  the  caseous 
parts,  but  they  are  also  found  in  the  tissue  generally,  and  often 
collected  in  large  cells.  In  size  they  vary  to  a  marked  extent.  In 
the  cells  they  often  form  compact  masses  of  short  bacilli,  but  in 
other  parts,  both  in  collections  and  singly,  they  attain  a  greater 
length  than  is  observed  in  any  other  form  of  tuberculosis.  Some  of 
the  bacilli  present  a  vary  interesting  appearance.  They  are  provided 

26 


402  •      INFECTIVE    DISEASES. 

terminally  with  a  sharply  defined  ovoid  body.  There  are  also 
collections  of  short  bacilli,  many  with  these  spore-like  appearances. 
The  author  has  also  seen  free  ovoid  forms,  sometimes  singly,  some- 
times in  groups.  From  their  connection  with  the  bacilli  and  their 
sharply  defined  outline  they  are  very  suggestive  of  spores. 

Johne  examined  the  livers  of  a  number  of  fowls  accidentally 
infected  by  phthisical  sputum.  Nocard  reported  an  outbreak  in 
a  poultry-yard  where  the  man  in  charge  had  consumption.  He 
also  found  the  disease  amongst  fowls  fed  with  the  infected  organs 
of  tubercular  cattle.  Subcutaneous  inoculation,  and  feeding  of  fowls 
with  sputum  or  bovine  virus,  will  produce  the  disease. 

Experimental  inoculation  of  tubercular  virus  from  different 
sources  affords  an  illustration  of  the  different  pathogenic  effects 
obtained  by  varieties  of  the  same  species  of  bacillus.  The  bacillus 
of  fowl- tuberculosis  is  a  distinct  variety.  A  very  small  proportion 
of  guinea-pigs,  inoculated  in  the  peritoneal  cavity  with  fowl-tubercle, 
succumb  to  the  disease,  though  so  susceptible  to  the  effects  of  human 
or  bovine  virus.  Maffucci  maintains  that  guinea-pigs  have  an 
immunity,  and  that  rabbits  rarely  develop  a  generalised  tuberculosis. 
Cultures  are  not  identical  in  appearance  with  those  obtained  from 
man,  and  on  microscopical  examination  show  many  long,  thick,  and 
branched  forms,  which  are  only  rarely  found  in  cultures  from  a 
human  source. 

Stamping-out  System. — In  1888  a  Departmental  Committee 
was  appointed  to  inquire  into  pleuro-pneumonia  and  tuberculosis, 
and  they  considered  that  legislation  ought  to  be  directed  not  only 
to  the  protection  of  cattle  from  tuberculosis,  but  also  to  prevent 
the  possibility  of  the  disease  being  communicated  to  man. 

The  following  extracts  are  from  the  recommendations  of  the 
Committee,  which  were  made  on  the  lines  of : — 

A.  PREVENTION. 

B.  EXTIRPATION. 

A. — Preventive  Measures. 

These  should  include  provision  for  : — 

Improved  hygiene  of  cattle  sheds,  etc.  (especially  in  the  direction  of 
providing  proper  ventilation,  pure  water  supply,  and  adequate  disinfection 
of  stalls,  etc.,  wherein  tubercular  animals  have  been  kept).  This  has 
been  partly  met  in  the  Dairy  and  Milk  Shops  Order,  but  its  administra- 
tion by  the  local  health  authorities  is  at  present  imperfect  ;  and  we  would 
suggest  that  it  should  be  much  more  stringently  enforced,  and  that 
veterinary  inspectors  should  be  given  more  extended  powers  of  entry  into 
all  places  where  animals  are  kept. 


TUBERCULOSIS.  403 

Improvement  in  the  hygienic  surroundings  of  animals  should  include 
isolation  of  all  suspected  cases,  precautions  against  the  flesh  or  milk  of 
diseased  animals  being  given  as  food  to  others — e.g.,  to  pigs,  fowls,  etc. — 
and  care  that  fodder,  litter,  and  water  should  not  be  taken  from  one 
animal  or  stall  and  given  to  another. 

Our  attention  has  been  drawn  to  the  frequency  with  which  animals, 
obviously  diseased,  sometimes  even  in  the  last  stage  of  the  malady,  are 
sold  in  open  market. 

Although  in  England  and  Ireland,  under  the  provisions  of  the 
Nuisances  Removal  Act  as  embodied  in  the  Public  Health  Act,  1885, 
the  medical  officer  of  health  or  inspector  of  nuisances  may  seize  such 
animals,  yet  such  seizure  is  rarely  performed. 

We  find  the  veterinary  inspector  has  no  power  to  prevent  such  sales, 
or  to  seize  the  beasts  for  slaughter,  since  tuberculosis  is  not  included  in 
the  Contagious  Diseases  (Animals)  Act  of  1878. 

We  further  find  that  there  is  actually  a  regular  trade  in  such  stock 
infected  with  tuberculosis,  and  that  they  go  by  the  name  of  "  wasters  " 
and  "  mincers,"  being  frequently  slaughtered  in  the  neighbourhood  of  the 
larger  towns,  to  which  such  portions  of  the  meat  as  are  likely  to  escape 
the  observation  of  the  inspector  of  nuisances  are  sent,  for  the  purposes 
of  sale  among  the  poorer  inhabitants,  and  especially  for  the  making  of 
sausages. 

We  are,  therefore,  very  strongly  of  opinion  that  power  should  be  given 
to  the  veterinary  inspector  to  seize  all  such  animals  in  fairs,  markets,  or 
in  transit. 

Notwithstanding  the  uniform  prevalence  of  the  disease  in  Europe  and 
elsewhere,  there  seems  to  be  no  reason  to  apprehend  that,  with  our 
present  regulations  for  the  slaughter  of  animals  at  the  port  of  debarka- 
tion, and  for  quarantine  of  those  imported  for  breeding,  there  is  any 
special  danger  of  increasing  the  infection  in  England  by  introduction 
from  abroad.  The  danger,  however,  exists  in  regard  to  the  stock  brought 
from  countries,  which  are  exempt  from  slaughter  on  landing,  and  sub- 
jected to  the  ordinary  veterinary  inspection  during  the  present  period  of 
detention  of  twelve  hours. 

It  is,  therefore,  evident  that  the  present  rules  for  the  prevention  of 
the  introduction  of  disease  into  the  United  Kingdom  from  abroad,  are 
incomplete. 

Since  all  authorities  are  agreed  that  the  disease  is  very  marked  by 
heredity,  we  think  it  highly  desirable  that  breeders  should  in  their  own, 
as  well  as  in  the  public  interest,  discontinue  breeding  from  tuberculous 
stock. 

B. — Extirpation. 

In  order  to  insure  the  gradual  extirpation  of  tuberculosis,  we  are  of 
opinion  that  it  should  be  included  in  the  Contagious  Diseases  (Animals) 
Acts  for  the  purposes  of  certain  sections  of  those  Acts,  so  as  to  provide  : — 

(a)  For  the  slaughter  of  diseased  animals,  when  found  diseased  on 
the  owner's  premises. 


404  INFECTIVE   DISEASES. 

(6)  For  the   payment  of  compensation  for  the   slaughter   of    such 
animals. 

(c)  For  the  seizure  and  slaughter  of  diseased  animals  exposed  in  fairs, 

markets,  etc.,  and  during  transit. 

(d)  For  the  seizure  and  slaughter  of  diseased  foreign  animals  at  the 

place  of  landing  in  this  country. 

Notification  of  this  disease  should  not  be  compulsory,  because  it  may 
exist  without  developing  any  sufficient  outward  evidence  to  enable  the 
owner  to  detect  it,  and  its  growth  is  so  slow,  that  non-notification  of  its 
•existence,  even  in  a  large  number  of  cases,  would  do  little  to  nullify  the 
stamping-out  effect  of  the  Act  of  1878. 

The  powers  and  responsibilities  of  inspectors  in  ordering  the  slaughter 
of  diseased  animals  should  be  the  same  for  tuberculosis  as  for  pleuro- 
pneumonia,  according  to  section  51  (5)  of  the  Act  of  1878. 

Further,  tubercle,  though  hereditary,  is  nevertheless  much  less  conta- 
gious than  the  other  diseases  included  under  the  Act  of  1878,  and  it  is 
clear,  therefore,  that  the  immediate  slaughter  of  diseased  animals  would 
go  far  to  stamp  it  out,  though,  doubtless  owing  to  heredity,  this  stamping- 
out  process  would  be  gradual  in  its  effect. 

A  supplementary  report  was  made  by  Professor  Horsley,  in 
which  he  expressed  the  opinion  that  there  ought  to  be  legislation 
to  prevent  breeding  from  diseased  animals,  and  compulsory  notifi- 
cation : — 

1.  Breeding. 

Tuberculosis  is  notorious,  even  among  the  laity,  as  a  disease  which  is 
transmitted  from  parent  to  offspring.  This  is  a  fact  with  which  cattle 
breeders  are  specially  familiar,  and  which  finds  strong  expression  in  the 
evidence  attached  to  this  report.  Further,  this  generally  received  truth 
has  been  completely  confirmed  by  the  results  of  scientific  investigation, 
as  is  also  duly  set  f6rth  in  the  report.  Considering,  therefore,  the 
extreme  importance  of  this  point,  I  think  that  the  act  of  wittingly  breed- 
ing from  animals  so  affected  should  be  made  an  indictable  offence.  The 
only  objection  that  can  be  raised  to  such  legislation,  which  if  effected 
would  prevent  the  dissemination  of  the  disease  among  cattle  in  this 
country,  is  that,  owing  to  the  present  state  of  want  of  knowledge  among 
cattle  owners,  and  even  veterinary  surgeons,  of  the  early  symptoms,  and 
physical  signs  on  examination,  of  this  disease,  prosecutions  would  occa- 
sionally occur  in  cases  in  which  no  fault  could  properly  be  attributed  to 
the  owner,  and  that,  therefore,  such  prosecutions  would  be  needlessly 
vexatious. 

Considering,  however,  the  extreme  rarity  with  which  such  cases  would 
occur,  and  that,  as  in  the  matter  of  non-notification,  each  case  would  be 
tried  before  district  magistrates  on  its  own  merits,  this  objection  is 
•deprived  of  the  force  it  might  have  possessed. 


TUBERCULOSIS.  405 

2.  Notification  of  the  Existence  of  the  Disease. 

This  point' requires  no  explanation,  since  it  is  clear  that,  unless  the 
veterinary  inspectors  or  authorities  receive  information  of  occurrence  of 
diseases,  it  is  impossible  to  ensure  the  thorough  carrying  out  of  the 
provisions  of  the  Contagious  Diseases  (Animals)  Act. 

That  deliberate  non-notification  should  be  punished  cannot  be  doubted 
by  any  one.  Objection,  however,  to  legislation  in  this  direction  has  been 
put  forward,  on  the  same  grounds  as  those  upon  which  the  prevention  of 
breeding  from  diseased  animals  was  contested.  As,  however,  I  consider 
that  these  objections  have  been  already  shown  to  have  no  weight,  I 
recommend  that  both  the  forbiddance  of  breeding  from  diseased  animals, 
and  the  notification  of  the  disease,  should  be  included  in  any  legislation 
for  tuberculosis. 

The  difficulty  referred  to  by  the  Committee,  is  presented  by 
cases  of  the  disease  which  cannot  be  detected  by  the  ordinary 
methods  of  examination,  and  might  possibly  be  overcome  by  the 
use  of  tuberculin  as  a  diagnostic  agent. 


CHAPTER  XXIX. 

LEPROSY. — SYPHILIS. — RHINOSCLEROMA. — TRACHOMA. 

LEPROSY. 

LEPROSY  occurs  in  three  forms — tubercular,  anaesthetic,  and  mixed 
tubercular.  It  may  be  classed  with  the  granulomata,  as  the  most 
common  form  of  the  disease  is  characterised  by  deposits  in  the  skin, 
mucous  membrane,  and  internal  organs.  These  deposits  are  composed 
of  small  cells,  and  large  cells  resembling  giant  cells.  The  cells 
become  deposited  in  the  surrounding  tissues,  and  so  the  tubercle 
enlarges,  involving  the  epidermis  and  developing  into  an  ulcerating 
sore ;  or,  after  a  certain  stage  of  development,  beginning  to  decline, 
and  finally  leaving  a  puffy  discoloration.  In  the  ansesthetic  form 
the  cells  invade  the  connective  tissue  of  nerves.  In  the  mixed 
form  the  varieties  occur  together,  but  the  tubercular  character 
predominates. 

Tubercular  leprosy  commences  with  the  development  of  an 
erytheinatous  patch,  which  becomes  infiltrated,  and  finally  tubercu- 
lated,  the  tubercles  varying  in  size  from  a  millet  seed  to  a  marble, 
or  even  larger.  The  eruption  on  the  head  and  face  produces  a 
characteristic  leonine  expression.  The  progress  of  the  disease  is  very 
slow.  After  death  the  following  changes  may  be  found  in  the 
internal  organs  :  Cirrhosis  of  the  liver  and  spleen,  enlargement  of 
the  lymphatic  glands,  and  a  condition  of  the  lungs  corresponding 
to  cheesy  bronchial  pneumonia. 

In  the  ansesthetic  form  patches  develop  on  the  skin,  which 
become  anaesthetic ;  ulceration  follows,  and  the  fingers  and  toes,  or 
the  entire  hand  and  foot,  may  slough  off. 

The  disease  is  undoubtedly  communicable,  but  the  infectivity  is 
of  a  very  low  type. 

The  infectiousness  is  illustrated  by  the  well-known  case  of  Father 
Damien.  Arning  inoculated  a  man  named  Keanu,  a  condemned 
criminal,  and  leprosy  developed  three  years  afterwards,  but  this  case 

406 


LEPROSY.  407 

is  not  regarded  as  conclusive,  as  the  man  had  a  family  history  of 
leprosy.  The  disease  has  never  been  known  to  spread  from  patients 
in  this  country,  who  have  contracted  the  disease  abroad. 

The  bacilli  of  leprosy  were  first  observed  by  Hansen  in  1874,  and 
subsequently  fully  described  by  him,  and  his  observations  confirmed 
by  Neisser,  in  1879. 

Bacillus  Leprae. — Rods  5  to  6  /x  in  length  and  1  /x  in  breadth. 
The  bacilli  are  straight  or  curved,  resembling  very  closely  the 
tubercle  bacilli.  They  are  present  in  the  leprous  tubercles  of  the 
skin  and  mucous  membrane,  in  the  lymphatic  glands,  and  in  the 
liver,  testicles,  and  kidney;  and  in  the  nerves  in  the  anaesthetic 
variety.  They  are  found  between  the  cells,  and  in  colonies  in  the 
cells.  They  stain  readily  with  the  aniline  dyes,  especially  by  the 
Ziehl-Neelsen  and  Gram's  methods.  The  bacilli  are  found  in  extra- 
ordinary numbers  in  the  skin;  and  they  are  rather  straighter  than 
tubercle  bacilli,  and  stain  more  readily. 

Numerous  unsuccessful  attempts  to  cultivate  the  bacillus  have 
been  made  by  many  bacteriologists.  The  author  has  made  repeated 
inoculations  upon  glycerine-agar,  upon  which  the  tubercle  bacillus 
grew  abundantly,  but  always  with  disappointing  results.  On  the 
other  hand,  Bordoni-Uffreduzzi  showed  the  author  a  cultivation 
which  he  had  obtained  from  the  bone  marrow  of  a  leper.  The 
cultivation  was  made  on  blood  serum  and  glycerine,  and  cover- 
glass  preparations  resisted  decolorisation  with  acid.  There  were 
slight  morphological  differences  when  compared  with  the  -appear- 
ance of  bacillus  leprse  in  the  tissues,  and  the  results  were  hardly 
conclusive. 

The  English  Leprosy  Commission  also  reported  successful  cultiva- 
tion of  the  leprosy  bacillus.  The  author  had  the  opportunity  of 
examining  one  of  the  first  cultures  received  in  this  country,  and 
found  that  the  bacilli  stained  deeply  in  ordinary  cover-glass  pre- 
parations, they  did  not  resist  decolorisation  by  the  Ziehl-Neelsen 
method,  and  they  corresponded  in  culture  with  one  of  the  varieties 
of  Bacillus  subtilis,  commonly  found  on  the  skin. 

Inoculation  of  animals  has  given  equally  unsatisfactory  results. 
Numerous  experiments  have  been  made  by  Beaven  Rake  on  small 
animals  and  birds,  with  invariably  negative  results.  The  blood  of 
leprous  patients,  tubercles  from  the  living  subject,  fragments  of  the 
skin  and  of  the  internal  organs  after  death,  have  been  inoculated  by 
different  observers  without  result.  Melcher  and  Ortmann  alone  claim 
to  have  produced  really  definite  results.  These  observers  excised 
leprous  tubercles  from  the  living  subject,  a'nd  inoculated  fragments 


408  INFECTIVE   DISEASES. 

in  the  anterior  chamber  of  the  eye  of  rabbits.  The  animals  died 
after  some  months  with  extensive  deposits  in  the  ccecum,  lymphatic 
glands,  spleen,  and  lungs. 

These  tubercles  varied  in  size  from  a  pin's  head  to  a  millet  seed, 
and  contained  bacilli,  resembling  leprosy  bacilli  in  their  staining 
reactions.  The  question  naturally  arises  whether  the  lesions  were 
really  indicative  of  leprosy  or  tuberculosis.  Until  the  experi- 
ments are  independently  confirmed,  and  the  result  of  inoculation 
differentiated  from  tuberculosis,  it  would  be  rash  to  accept  these 
experiments  as  conclusive. 

It  has  been  suggested  that  tuberculosis  and  leprosy  are  identical. 
There  is  a  similarity  in  the  bacilli  and  in  the  lesions  of  leprosy  and 
tuberculosis,  the  injection  of  tuberculin  produces  a  reaction  in  leprosy 
nodules,  and  many  lepers  die  from  tubercular  disease  of  the  lung. 
But  while  tuberculosis  is  very  readily  transmitted  to  guinea-pigs 
and  rabbits  by  inoculation  of  fragments  of  tubercular  tissue,  leprosy 
is  inoculable,  if  at  all,  in  most  exceptional  instances.  The  bacilli 
of  tubercle  are  cultivated  with  the  greatest  facility,  the  bacilli  of 
leprosy,  if  at  all,  only  with  exceptional  difficulty;  tubercle  bacilli 
are  found  in  giant  cells,  leprosy  bacilli  in  the  so-called  leprosy 
cells.  Leprosy  bacilli  are  straighter  than  human  tubercle  bacillir 
and  differ  slightly  in  their  behaviour  to  staining  reagents.  On 
the  other  hand,  the  morphological  differences  are  not  greater  than 
those  existing  between  different  forms  of  tubercle  bacilli  obtained 
from  tuberculosis  in  animals  and  birds.  It  would  be  premature 
to  regard  leprosy  as  a  variety  of  tubercle  until  cultivations  of 
the  bacillus  have  been  obtained,  and  carefully  compared  with 
those  of  the  tubercle  bacillus.  Differences  in  morphological  details 
and  results  of  inoculation  would  then  carry  less  weight  as  a  means 
of  differentiation. 

The  tubercular  pneumonia  of  lepers  would  be  regarded,  if  the 
bacilli  are  identical,  as  a  development  of  leprosy  in  the  lungs,  and 
not,  as  at  present,  a  result  of  double  infection  with  tuberculosis. 

METHODS  OF  EXAMINING  THE  BACILLUS  OF  LEPROSY. 

Cover-glass  preparations  may  be  made  in  the  ordinary  way,  or  by  a 
special  method,  which  consists  in  clamping  a  nodule  with  a  pile  clarnp 
until  a  state  of  anaemia  of  the  tissue  is  produced.  On  pricking  with  a 
needle  or  sharp  knife  a  drop  of  clear  liquid  exudes,  from  which  cover- 
glass  preparations  may  be  made,  and  stained  by  Neelsen's  method. 

For  sections  the  author  prefers  Neelsen's  method  and  methylene-blue. 
They  can  also  be  stained  by  Gram's  method,  which,  as  a  rule,  brings  out 
very  clearly  the  beaded  appearance  of  the  bacilli. 


DESCRIPTION    OF    PLATE    XIY. 
Bacillus  Leprae. 

FIG.  1. — From  a  section  of  the  skin  of  a  leper.  The  section  is,  almost  in 
its  entirety,  stained  red,  and,  with  moderate  amplification,  has  a  finely 
granular  appearance.  Stained  by  the  Ziehl-Neelsen  method  (carbolised 
fuchsine  and  methylene-blue).  x  200. 

FIG.  2. — Part  of  the  same  preparation  with  high  amplification,  showing  that 
the  appearances  described  above  are  due  entirely  to  an  invasion  of  the 
tissue  by  the  bacilli  of  leprosy,  x  1500. 


; 


-        ,        :  -.- 

J 


Plgl. 


Tig  2 


BACILLUS  LEPTLE 


^SrtAtfp 

OF  THB 

UNIVERSITY 
Of 


LEPROSY.  409 

Method  of  Babes. — Preparations  are  stained  in  rosaniline  hydrochlorate 
in  aniline  water,  decolorised  in  33  per  cent,  hydrochloric  acid,  and  after- 
stained  with  methylene-blue. 

Stamping-out  System. — The  history  of  leprosy  in  the  British 
Islands  during  the  Middle  Ages,  and  the  conditions  under  which  it 
both  increased  and  declined,  have  been  discussed  by  several  writers. 
A  large  number  of  institutions  of  a  charitable  and  ecclesiastical 
character  were  established  in  endemic  areas  and  were  occupied  by 
the  lepers  either  voluntarily,  or  compulsorily  by  means  of  the  Act 
De  leproso  amovendo.  These  institutions  were  to  a  very  small  extent 
a  means  of  segregation.  According  to  Dr.  Newman  the  disease, 
which  had  reached  its  zenith  about  the  twelfth  or  thirteenth  century, 
began  to  decline  from  that  time  owing  to  "  a  general  and  extensive 
social  improvement  in  the  life  of  the  people,  to  a  complete  change 
in  the  poor  and  insufficient  diet  (which  it  is  evident  consisted  far 
too  largely  of  bad  meat,  salt,  putrid  and  dried  fish,  and  an  almost 
entire  lack  of  vegetables)  and  to  agricultural  advancement,  improved 
sanitation  and  land  drainage."  Of  all  the  unfavourable  conditions 
it  would  appear  that  food  in  some  way  was  especially  associated  with 
the  cause  of  the  disease,  either  by  introducing  the  bacillus  or  by 
rendering  the  tissues  a  suitable  soil  for  its  reception  and  development. 

In  other  countries  segregation  has  been  attempted  voluntarily 
or  compulsorily,  but  it  has  never  been  completely  carried  out.  There 
can  be  very  little  doubt  that  the  presence  of  a  leper  in  a  healthy 
community  is  no  greater  source  of  danger  than  the  presence  of 
an  individual  suffering  from  tuberculosis,  but,  for  other  reasons, 
voluntary  isolation  should  be  carried  out  as  completely  as  local 
circumstances  will  permit. 

The  Leprosy  Commission  in  India  recommended — 

(a)  That  the  sale  of  articles  of  food  and  drink  by  lepers  should 

be  prohibited,  and  that  lepers  should  be  prevented  from 
following  certain  specified  occupations. 

(b)  That  the  concentration  of  lepers  in  towns  should  be  discouraged. 

(c)  That  Leper  Asylums  should  be  established  in  which  lepers 

might  live  voluntarily. 

(d)  That  Leper  Farms   scattered   over   the   country  should   be 

encouraged. 

(e)  That  the  few  children  who   are  born  of   lepers  should   be 

removed  to  Orphanages. 

They  concluded  that  by  means  of   improved  sanitation  and   good 
dietetic  conditions  a  diminution  of  leprosy  will  result. 


410 


INFECTIVE   DISEASES. 


© 
® 


%1 


SYPHILIS. 

Syphilis  is  a  disease  peculiar  to  man,  and  communicable  'only  by 
inoculation.  The  local  infection  is  followed  by  a  period  of  latency, 
and  by  a  period  during  which  generalised  eruptions  appear.  One 
attack  confers  immunity  from  future  attacks.  The  virus  in  its 
most  virulent  form  is  found  in  the  primary  seat  of  inoculation,  and 

in  the  indurated  glands  which 
follow.  It  is  also  supposed  to  be 
present  in  the  blood  and  secretions. 
Lustgarten,  Eve  and  Lingard  have 
found  bacteria  which  they  believed 
to  be  specific. 

Bacillus  in  Syphilis  (Lustgar- 
ten).— Rods  resembling  the  bacilli 
of  leprosy  and  tuberculosis,  3  to  4  ^ 
long,  -8  //,  thick.  Two  or  more 
colourless,  ovoid  points  in  the  course 
of  the  rod  are  visible  with  a  high 

power ;  it  is  thought  that  possibly  they  are  spores.  The  bacilli 
are  always  found  in  the  interior  of  nucleated  cells,  which  are 
more  than  double  the  size  of 

leucocytes.      They    have    been   ob-  .   „.  ..^w.,**   .  

served  in  the  discharge  of  the  fc'fij^ 
primary  lesion,  and  in  tertiary  «V-&.^: 
gummata.  f ?$*$/'• 

Alvarez  and  Tavel  state  that 
an  identical  bacillus  is  found  in  '^l^v;; 
normal  secretions  (smegma).  Eve 
and  Lingard  have  described  a  bacil- 
lus associated  with  specific  lesions, 
which  differs  from  the  above  in  its 

morphology  and  behaviour  towards  FIG.  176.— WANDERING  CELL  CON- 
staining  reagents.  TAINING  BACILLI 


^  rim*  ^a&'viWdsssww*'*' 

FIG.  175.  —  COVER-GLASS  PREPARA- 
TION OF  PUS  FROM  A  CHANCRE,    X 

1050  (LUSTGARTEN). 


•:  #&, 


METHODS  OF  STAINING  THE  BACILLUS  OF  SYPHILIS. 

Method  of  Lustgarten : — 

Sections  are  placed  for  twelve  to  twenty-four  hours  in  the  following 
solution,  at  the  ordinary  temperature  of  the  room,  and  finally  the  solution 
is  warmed  for  two  hours  at  60°  C.  : — 

Concentrated  alcoholic  solution  of  gentian-violet      .  11 

Aniline  water     .  .....         100 


SYPHILIS.  411 

The  sections  are  then  placed  for  a  few  minutes  in  absolute  alcohol, 
and  from  this  transferred  to  a  1-5  per  cent,  solution  of  permanganate  of 
potash.  After  ten  minutes  they  are  immersed  for  a  moment  in  a  pure 
concentrated  solution  of  sulphurous  acid.  If  the  section  is  not  completely 
decolorised,  immersion  in  the  alcohol  and  in  the  acid  bath  must  be 
repeated  three  or  four  times.  The  sections  are  finally  dehydrated  with 
absolute  alcohol,  cleared  with  clove-oil,  and  mounted  in  Canada  balsam. 

By  this  method  the  bacillus  is  distinguished  from  many  bacteria,  but 
not  from  the  bacilli  of  tubercle  and  leprosy  which  are  stained  by  this 
process. 

Method  of  De  Giacomi : — 

Cover-glass  preparations  are  stained  with  hot  solution  of  fuchsine 
containing  a  few  drops  of  perchloride  of  iron.  They  are  then  decolorised 
in  strong  perchloride  of  iron,  and  after-stained  with  vesuvin  or  Bismarck- 
brown. 

Method  of  Doutrelepont  and  Schiltz  : — 

Sections  are  stained  in  a  weak  aqueous  solution  of  gentian-violet  and 
after-stained  with  safraniu. 

The  nature  of  the  contagium  in  syphilis  is  unknown. 

Protective  Inoculation. — Inoculation  of  the  virus,  or  syphilisa- 
tion,  as  a  protective  measure,  was  at  one  time  practised  and  strongly 
advocated ;  but  it  is  rightly  regarded  in  this  country  as  dangerous 
and  unjustifiable.  From  the  experiments  of  Bicord  it  would  appear 
that  the  local  results  in  the  vesicular  stage  resemble  the  results  of 
the  inoculation  of  virulent  vaccinogenic  grease  or  horse- pox.  The 
inoculation  goes  through  the  stages  of  papule,  vesicle,  ulcer,  scab, 
and  scar.  The  accidental  inoculation  which  occurs  in  cases  of 
vaccino-syphilis  may  so  closely  resemble  the  results  of  inoculation 
with  very  virulent  cow-pox,  that  it  is  sometimes  difficult  to  decide 
as  to  the  exact  nature  of  these  cases. 

RHIXOSCLEROMA. 

Rhinoscleroina  is  a  rare  disease,  resembling  lupus,  and  pro- 
ducing in  the  nostrils  and  neighbouring  parts  nodular  swellings, 
composed  of  granulation-tissue.  The  disease  is  met  with  in 
America,  Egypt,  Austria,  and  Italy.  There  are  no  giant  cells,  but 
peculiar  large  cells,  which  were  first  described  by  Mikulicz.  Frisch 
discovered  bacteria  in  sections,  and  Cornil  and  Alvarez  pointed  out 
the  existence  of  a  capsule.  In  morphology  and  cultivation  they 
resemble,  according  to  Dittrich,  Friedlander's  pneumococcus.  They 
are  probably  identical  with  this  micro-organism,  and  Paltauf  and 
Eiselsberg,  and  others,  found  that  they  produced  septicaemia  in  rabbits 
and  guinea-pigs. 

Bacterium   of   Rhinoscleroma   (Bcudttus  of  Rhinosderama, 


412  INFECTIVE   DISEASES. 

Cornil  and  Alvarez). — Cocci  and  short  rods,  1*5  to  3  /x,  in  length,. 
*5  to  '8  JJL  thick.  Deeply  coloured  points  or  granules  may  occur 
in  the  course  of  the  rods  when  stained,  but  it  is  very  doubtful 
whether  these  can  be  considered  as  spores.  The  bacteria  are  en- 
capsuled,  the  capsule  being  round  when  enclosing  a  coccus,  and 
ovoid  when  enclosing  a  rod.  The  capsule  is  composed  of  a  tough 
resisting  substance;  two  or  more  capsules  may  unite  by  fusion, 
enclosing  two,  three,  or  a  greater  number  of  rods.  The  bacilli  were 
observed  in  sections  of  the  tumours,  which  developed  on  the  lips 
and  in  the  nasal  and  pharyngo-laryngeal  regions. 

METHOD  OF  STAINING  THE  BACILLUS  OF  RHINO- SCLEROMA. 

Method  of  Cornil  and  Alvarez  : — 

Sections  are  immersed  in  a  solution  of  methyl-violet  (B)  for  twenty- 
four  to  forty-eight  hours,  with  or  without  the  addition  of  aniline-water  ; 
and  are  then  decolorised  after  treatment  with  the  solution  of.  iodine  in 
iodide  of  potassium.  If  the  sections  are  left  to  decolorise  in  alcohol  for 
forty-eight  hours,  the  capsule  is  rendered  visible. 

TRACHOMA. 

Trachoma  is  a  disease  of  the  conjunctiva,  common  in  Egypt. 
The  new  growth  is  composed  of  round  cells,  and  may  be  regarded, 
according  to  Kartulis,  as  the  chronic  stage  of  either  gonorrhoeal  or 
Egyptian  ophthalmia.  Koch  failed  to  find  any  micro- organisms 
in  the  swollen  lymph  follicles.  Sattler  asserted  that  he  had  culti- 
vated a  micrococcus  which  produced  the  disease  when  inoculated 
on  the  conjunctiva.  Other  observers  have  found  the  common 
pyogenic  micrococci  in  the  secretions,  especially  Staphylococcus 
pyogenes  aureus  and  albus. 


CHAPTER   XXX. 

ACTINOMYCOSIS. — MADURA   DISEASE. 
ACTINOMYCOSIS. 

ACTINOMYCOSIS  belongs  to  the  class  of  infective  granulomata.  It  is 
a  chronic  inflammatory  affection  characterised  by  the  presence  of  a 
special  microphyte,  which  by  irritation  produces  a  neoplasm,  composed 
-of  round  cells,  epithelioid  cells,  giant  cells,  and  fibrous  tissue.  These 
neoplasms  form  nodular  tumours  of  various  sizes.  In  some  cases 
there  is  a  tendency  to  develop  very  large  tumours,  and  in  others  to 
break  down  early  and  suppurate.  In  cattle,  cretification  takes 
place  in  the  fungus  tufts.  Actinomycosis  closely  resembles  tuber- 
culosis in  its  histological  characters.  The  disease  attacks  man, 
horses,  cattle,  and  pigs. 

Many  interesting  observations  have  been  made  upon  the  origin 
of  this  disease  in  man.  Two  cases  have  been  recorded  in  support  of 
the  theory  of  direct  infection  from  the  cow.  Stelzner  described  a 
case  of  actinomycosis  in  a  man  who  had  had  the  care  of  animals, 
some  of  which  had  suppurating  glands.  Hacker  had  a  case  of 
actinomycosis  of  the  tongue  in  a  man  who  had  charge  of  cows,  one 
of  which  had  a  tumour  of  the  jaw  which  he  had  opened.  On  the 
other  hand,  Moosbrugger  found  that  out  of  75  cases,  54  were  in  men, 
and  21  in  women,  including  2  children.  In  11  of  these  men  the 
occupation  was  not  stated.  In  33  their  occupation  did  not  bring 
them  into  contact  with  diseased  animals ;  they  were,  for  example, 
millers,  glaziers,  tailors,  shop  people,  and  students.  Only  10  cases 
occurred  among  farmers,  peasants,  and  farm-labourers,  and  in  only 
one  case  out  of  the  10,  had  the  patient  been  brought  into  contact 
with  diseased  animals. 

Out  of  the  21  women,  there  were  only  4  peasants,  and  none 
of  them  had  been  associated  with  diseased  cattle. 

Infection  by  the  flesh  of  diseased  animals  has  also  been  dis- 
cussed. But  there  is  no  evidence  of  prevalence  of  the  disease 

413 


414  INFECTIVE   DISEASES. 

among  slaughterers  and  butchers,  who  would  be  particularly  liable 
to  it,  if  flesh  were  a  source  of  infection.  The  chances  of  infection 
by  ingestion  are  minimised  by  the  flesh  being  almost  always  cooked. 
Actinomycosis  occurs  also  in  pigs,  and  pork  is  very  often  eaten  in  an 
uncooked  state  ;  but  Israel  has  pointed  out  that  this  may  probably 
be  excluded,  as  many  of  the  cases  occurred  among  strict  Jews. 

The  evidence  points  to  the  disease  originating  in  man  and  lower 
animals  from  the  same  source,  and  there  is  a  very  strong  suspicion 
attached  to  cereals.  This  view  is  supported  by  important  obser- 
vations, with  reference  to  the  part  played  by  cereals  in  inducing 
the  disease  in  cattle,  and  it  gains  additional  support  from  a  case 
described  by  Soltmann,  where  the  disease  resulted  from  an  awn  of 
wall  barley.  A  boy,  aged  eleven,  accidentally  swallowed  an  awn 
of  Hordeum  murinum.  He  became  very  ill,  and  suffered  great  pain 
behind  the  sternum,  extending  to  the  back.  An  abscess  formed, 
covering  an  area  extending  over  six  intercostal  spaces,  and  when 
opened,  the  awn  of  this  grass  was  found  in  the  evacuated  pus. 
The  pain,  however,  continued,  and  fresh  deposits  occurred,  and  when 
the  boy  was  taken  to  the  hospital,  the  ray-fungus  was  detected. 
Possibly  the  spores  of  the  fungus  can  be  conveyed  both  by  air  and 
water. 

This  disease  in  cattle  has  long  been  known  in  this  country,  but 
its  various  manifestations  were  either  mistaken  for  other  diseases, 
or  simply  received  popular  names.  Indeed,  the  various  forms  are 
still  familiar  to  many  as  wens,  clyers  or  crewels,  scrofulous,  tuber- 
cular or  strumous  abscesses,  polypus,  lymphoma,  cancer  of  the 
tongue,  scirrhous  tongue,  indurated  tongue,  ulcerated  tongue,  cancer 
of  bone,  bone  tubercle,  osteo-sarcoma,  fibroplastic  degeneration  of 
bone,  spina  ventosa,  and  carcinoma. 

Bovine  antinomycosis  is  especially  prevalent  in  river  valleys, 
marshes,  and  on  land  reclaimed  from  the  sea.  The  disease  occurs 
at  all  times  of  the  year,  but  general  experience  leads  to  the  belief 
that  it  occurs  more  commonly  in  the  winter. 

It  is  more  frequently  met  with  in  young  animals,  and  usually 
occurs  between  one  and  three  years,  but  it  may  be  found  at  almost 
any  age,  and  probably  affects  equally  both  sexes. 

There  is  little  if  any  evidence  to  show  that  the  disease  is  heredi- 
tary. In  numerous  cases,  the  family  history  has  been  most  carefully 
inquired  into  by  the  author ;  and  in  the  case  of  some  imported 
pedigree  animals,  the  disease  was  quite  unknown  on  the  farm  where 
they  had  been  bred. 

The  tongue  is  so  commonly  the  seat  of  the  disease,  that  suspicion 


ACTINOM  YCOSIS.  415 

at  once  falls  on  food  as  the  means  by  which  the  parasite  is  conveyed. 
Skin  wounds  produced  by  rubbing  against  the  mangers,  posts,  or 
wire  fencing,  may  also  become  infected. 

The  evidence  is  very  strong  in  favour  of  believing  that  the  micro- 
organism gains  access  to  the  system  through  wounds  or  lacerations 
of  the  mucous  membrane  and  skin,  or  through  carious  teeth.  It 
has  been  pointed  out  that  the  common  occurrence  of  the  disease 
at  the  time  of  the  second  dentition  may  be  owing  to  the  wounds 
produced  in  the  alveolar  mucous  membrane  by  the  shedding  of 
the  teeth.  Experience  also  points  to  straw  being  sometimes  a 
factor  in  the  production  of  the  disease,  and  it  is  possible  that  thistles 
and  frozen  roots  also,  by  wounding  the  mucous  membrane,  may 
afford  a  way  for  the  entrance  of  the  micro-organism.  The  disease 
in  the  jaws,  both  in  man  and  in  cattle,  is  very  commonly  associated 
with  carious  teeth. 

The  cowsheds,  pastures,  and  drinking  tanks  may  become  infected 
with  the  discharges  from  diseased  animals.  The  discharge  con- 
taminates the  fodder  in  the  sheds,  and  falls  on  thistles  and  siliceous 
grasses  in  the  pasture,  which  may  first  wound,  and  then  introduce 
t]je  micro-organism.  The  discharge  is  also  coughed  out  of  the 
mouth,  and  expelled  from  the  nose,  in  cases  in  which  a  tumour  in 
the  pharynx,  or  the  nasal  chambers,  has  undergone  suppuration. 

Jensen  believed  that  the  disease  was  produced  by  different  kinds 
of  grain,  especially  when  cultivated  on  ground  reclaimed  from  the 
sea.  He  mentions  an  instance  of  a  farm,  where  nearly  the  whole 
of  the  young  stock,  about  thirty  in  number,  had  actinomycosis  after 
feeding  on  mixed  forage,  grown  on  a  certain  field.  Two  years  after- 
wards the  same  disease  occurred  in  the  same  stalls  in  four  animals, 
after  being  fed  on  barley-straw  from  the  same  field.  According  to 
Jensen,  the  fungus  grows  on  grain,  husks,  and  straw  of  different 
cereals,  but  most  abundantly  on  barley,  which  is  also  the  most 
likely  to  wound  the  mucous  membrane.  Johne's  observations  tend 
to  corroborate  this  view,  for  in  twenty-two  out  of  twenty-four  cases 
in  which  he  found  barley  sticking  in  the  tonsils  of  pigs,  he  found 
the  beard  thickly  beset  with  a  fungus  very  similar  to,  if  not  identical 
with,  the  ray-fungus.  These  observations  are  of  great  interest  in 
connection  with  Soltmann's  case. 

Experience  points  to  the  belief  that  the  disease  is  not  readily 
communicable  from  animal  to  animal,  and  it  is  possible  that  when 
it  affects  a  large  number  of  cattle  in  a  herd,  the  same  causes  have 
been  acting  to  produce  the  disease  in  a  number,  which  in  another 
in.-tunce  may  only  produce  it  in  one.  At  the  same  time,  isolated 


416  INFECTIVE   DISEASES. 

cases -are  possibly  not  quite  so  common  as  they  are  reported  to  be. 
It  is  well  known  that,  as  a  rule,  the  services  of  a  veterinary  surgeon 
are  not  called  for  except  in  hopeless  or  very  severe  cases.  The 
cowmen  themselves,  in  many  districts,  treat  the  cows  successfully, 
and  then  send  them  into  the  market,  and  thus  the  existence  of 
previous  cases  may  not  have  come  to  the  knowledge  of  the  veterinary 
surgeon. 

Historical. — In  1845  Professor  von  Langenbeck,  of  Kiel,  made 
notes  of  a  case  of  vertebral  caries  in  a  man,  and  prepared  drawings 
of  peculiar  bodies  in  the  pus  from  an  abscess.  The  drawings  were 
published  together  with  a  reference  to  the  case  by  Israel  in  1878. 
There  can  be  little  doubt  that  these  structures  were  the  fungi  of 
actinomycosis.  But  the  first  to  publish  observations  was  Lebert 
in  1848. 

Lebert  received  from  M.  Louis  some  pus,  of  a  thick,  almost 
gelatinous  consistency,  which  had  been  obtained  from  an  abscess  of 
the  thoracic  wall  in  a  man  aged  fifty.  The  patient  had  been  attacked 
four  months  previously  by  a  pulmonary  affection,  which  was 
suspected  by  M.  Louis  to  be  cancerous  in  nature.  The  pus  contained 
a  very  considerable  number  of  little  spherical  bodies  of  a  slightly 
greenish-yellow  colour,  about  the  size  of  a  pin's  head.  They 
could  be  readily  crushed  between  two  strips  of  glass,  and  on 
examination  with  a  power  of  fifty  diameters  two  elements  could  be 
distinguished  :  a  soft  connective  substance,  and  many  hard,  narrow, 
wedge-shaped  corpuscles,  arranged  in  a  radiating  manner.  Under 
a  high  power  these  bodies  were  observed  to  be  -2^  to  ^  of  an  inch 
in  length,  -^^  in  width  at  the  base,  and  -.^  in  width  at  the 
apex.  Some  of  these  corpuscles  were  regular,  while  others  showed 
one  or  two  constrictions,  with  intermediate  flask-shaped  swellings. 
Lebert  tested  these  structures  with  reagents,  with  the  following 
results.  The  bodies  were  found  to  remain  unaltered  by  concentrated 
mineral  acids.  Acetic  acid  freed  them  from  foreign  elements 
adhering  to  their  surface.  Solution  of  caustic  potash  did  not  affect 
them  if  used  cold,  but  a  boiling  solution  reduced  the  cuneiform 
structures  to  a  fine  greyish  powder  without  dissolving  them.  Ether, 
alcohol,  and  chloroform  had  no  effect  upon  them  when  used  either 
hot  or  cold.  Solution  of  potash,  in  which  these  bodies  had  been 
heated,  mixed  with  a  solution  of  sulphate  of  copper  and  brought  to 
boiling  point,  did  not  offer  any  uniform  red  colour,  which  would 
have  been  the  case  if  they  had  contained  albumin.  Thus,  the  chief 
chemical  characters  of  albuminous  and  fatty  substances  were 
wanting,  and  they  resembled  chitine  in  their  behaviour  to  reagents. 


ACTINOMYCOSIS. 


41  r 


Lebert  bore  in  mind  the  possible  existence  of  some  helminthic  debris, 
of  which  these  bodies  might  be  booklets,  but  he  sought  in  vain  for 
echinococci  and  cysticerci. 

Actinomycotic  pus  was  later  described  and  figured  by   Robin. 
In  the  illustration  accompanying  the  description,  the  fungi  are  most 


FIG.  177. — SECTION  OF  LIVER  FROM  A  CASE  OF  ACTINOMYCOSIS  IN  MAX. 

accurately  depicted.  Robin  states  that  he  had  found,  in  two  or 
three  cases  in  the  pus  of  deep-seated  chronic  abscesses,  yellowish  grains 
attaining  a  diameter  of  one-tenth  of  a  mm.,  surrounded  by  a  sort  of 
halo  or  thin,  viscous,  finely  granular  stratum,  containing  leucocytes. 
These  grains  were  composed  of  elements  2  to  6  mm.  in  length,  swollen 

27 


418  INFECTIVE    DISEASES. 

at  one  end  and  tapering  off  at  the  other,  arranged  in  a  regular  series, 
radiating  from  a  common  centre  which  consisted  of  granular  matter. 
They  were  highly  refractive,  possessed  a  brilliant  centre  and  sharply 
-defined  outline  ;  they  were  dissolved,  or  at  least  rendered  indistinct, 
by  acetic  acid,  and  proved  insoluble  in  ammonia  and  ether. 

The  disease  in  man  was  next  described  by  Israel  in  the  paper 
mentioned  above.  Ponfick  was  the  first  to  clearly  recognise  the 
identity  of  the  disease  in  man  with  the  disease  in  cattle,  and  he 
described  a  number  of  cases  in  man.  Israel  subsequently  published 
a  work  on  the  subject.  The  various  cases  which  had  been  observed 
up  to  that  date  were  described,  and  the  disease  classified  according 
to  the  seat  of  invasion. 

From  this  time  onwards  numbers  of  cases  in  man  have  been 
described,  and  various  important  researches  published,  of  which 
those  of  Bostrom  and  Moosbrugger  may  be  especially  mentioned. 

In  England,  Acland  recognised  a  case  on  examining  the  liver 
after  death  (Fig.  177).  H.  Taylor  was  the  first,  in  this  country, 
to  detect  the  fungus  during  the  life  of  a  patient.  Shattock  found 
.specimens  of  the  disease  in  museums.  Skerrit,  Powell  and  Godlee, 
Eve,  Delepine,  Hansome,  Poore,  Malcolm  Morris  and  others  have 
published  cases. 

In  Italy,  Perroncito  studied  the  sarcomata  of  cattle,  and  claims 
to  have  first  observed  the  micro-organism  in  1863.  In  1875  he 
described  it  in  the  Encyclopaedia  Agraria,  and,  from  the  negative 
results  obtained  by  inoculation  experiments,  was  led  to  regard  it, 
not  as  the  cause,  but  as  a  result  of  the  disease. 

Rivolta  of  Turin  also  claims  to  have  been  the  first  to  have 
•discovered  the  fungus  in  actinomycosis  bovis.  As  early  as  1868 
he  published  a  paper  on  a  sarcoma tous  tumour  of  the  jaw  of 
-an  ox. 

Hahn  of  Munich,  in  1870,  undoubtedly  met  with  the  fungus,  for 
he  states  that  in  a  case  of  "  wooden-tongue  "  he  found  characteristic 
organised  structures,  which  he  provisionally  described  as  a  species  of 
mould  fungus. 

Bellinger  was  the  first  to  recognise  the  nature  of  this  disease  in 
cattle.  In  1876  he  pointed  out  that  new  growths  occasionally 
occurred  on  the  upper  and  lower  jaws  of  cattle,  which  either  started 
from  the  alveoli  of  the  back  teeth,  or  from  the  spongy  tissue  of  the 
bone,  and  by  increasing  in  size  loosened  the  teeth.  In  their  progress 
they  destroyed  bone,  muscles,  mucous  membrane,  and  skin.  After 
some  time  they  frequently  broke  down,  forming  ulcers,  abscesses, 
and  fistulas  ;  but  in  some  cases  tumours  were  formed,  which  attained 


ACTINOMYCOSIS.  419 

the  size  of  a  child's  head.  Bellinger  stated  that  this  disease  had 
been  known  by  various  names, — Osteosarkome,  Winddorn  (Spina 
ventosa),  Knochenkrebs,  Knochenwurm  ;  in  other  instances  it  had  been 
regarded  as  bone  tuberculosis,  or  mistaken  for  a  simple  chronic 
glossitis.  Among  breeders  of  cattle  and  owners  of  stock  in  Germany 
it  had  been  known  under  the  following  names  :  Ladendruck,  Laden- 
geschwulst,  dicker  Backen,  £dckel,  Kinnbeule,  Kiefergeschwulst,  etc. 

Bollinger  pointed  out  that  these  swellings  consisted  of  several 
centres  of  growth,  bound  together  by  connective  tissue.  They  were 
often  as  large  as  a  walnut  or  a  hen's  egg,  and  of  a  pale  yellow 
colour  and  moist  appearance.  The  cut  surface  presented  yellowish- 
white,  suppurative  foci,  while  in  other  cases  the  growths  had  a 
spongy  texture,  owing  to  the  formation  of  lacunae  or  hollow  spaces 
in  a  fibrous  stroma,  which  contained  a  turbid,  thick,  yellow,  caseous 
pulp. 

Microscopical  examination  of  the  tumour  showed  that  it  had  a 
structure  like  a  sarcoma,  while  the  squeezed-out  pulp  consisted 
principally  of  pus  cells,  granulation  cells,  fat  granules,  and  granular 
detritus.  In  addition,  there  were  numerous  opaque,  pale-yellow,  and 
coarsely  granular  bodies  of  different  sizes,  which  had  a  mulberry- 
like  appearance,  and  were  sometimes  encrusted  with  chalk.  After 
careful  examination  Bollinger  found  that  these  bodies  were  true 
fungi,  and  he  further  maintained,  from  the  constancy  of  their 
appearance  in  all  parts  of  the  sarcomatous  growth,  that  they  were 
not  accidental,  but  of  pathogenic  significance.  This  was  found  to 
be  the  case,  not  only  in  fresh  preparations,  but  in  old  specimens 
preserved  in  the  museum.  This  remarkable  form  of  mycosis  was 
observed  by  Bollinger,  not  only  in  the  upper  and  lower  jaws,  but 
also  in  the  tongue.  It  had  long  been  observed  that  the  tongue  was 
sometimes  covered  with  more  or  less  tubercular  growths,  scattered 
abundantly  over  the  surface  of  the  mucous  membrane,  mostly  the  size 
of  a  millet  seed  or  hemp  seed,  but  often  reaching  the  size  of  a  cherry 
or  walnut,  or  even  larger.  In  the  fresh  state  these  nodules  were 
greyish -white,  and  semi- transparent,  but  they  soon  became  cloudy 
or  distinctly  puriform  in  the  centre ;  they  were  surrounded  externally 
with  a  connective  tissue  capsule.  If  the  nodules  were  situated  on 
the  surface  of  the  tongue,  destruction  of  the  mucous  membrane  very 
readily  followed,  leading  to  the  formation  of  ulcers.  The  tongue 
also  might  become  affected  with  an  interstitial  glossitis,  which  often, 
in  spite  of  the  partial  atrophy  of  the  muscular  fibres,  led  to  a  great 
enlargement  and  wood-like  hardness  of  the  tongue.  On  account 
of  this  peculiar  character,  such  a  tongue  was  long  known  in  South 


420  INFECTIVE   DISEASES. 

Germany  as  Holzzunge.  In  other  cases  the  condition  was  regarded 
as  "  tubercle  of  the  tongue,"  "  chronic  sarcoma,"  "  chronic  interstitial 
glossitis,"  or  simply  "  degeneration  of  the  tongue." 

Bollinger  described  this  disease  as  occurring  in  cattle  of  all  agesy 
developing  itself  gradually,  and  being  always  incurable.  As  a  ruler 
the  animals  were  slaughtered,  because  the  diminished  mobility  and 
enlargement  of  the  tongue  interfered  with  feeding.  He  also  pointed 
out  that  this  disease  of  the  tongue  was  by  no  means  rare,  as  he  had 
had  no  less  than  six  such  tongues  from  different  parts  of  Bavaria  in 
the  space  of  a  year,  and  he  also  had  been  able  to  prove  the  existence 
of  the  disease  in  museum  specimens. 

On  further  continuing  his  researches,  Bollinger  found  the  same 
fungus  in  tumours  which  occurred  in  the  pharynx,  larynx,  and 
the  mucous  membrane  of  the  stomach.  These  tumours  were  very 
common  in  the  throat  in  some  parts  of  North  Germany,  where  as 
many  as  5  per  cent,  of  the  animals  had  been  known  to  be  affected. 
The  disease  frequently  occurred  in  the  form  of  subcutaneous 
neoplasms,  called  Lymphome,  Hohzgeschivulste,  Fibrome,  Tuberkel, 
Tuberkel-scropheln. 

This  disease  also  appeared  in  the  form  of  abscesses,  which  were 
called,  in  many  districts,  Schlundbeulen.  These  growths  were  found 
in  the  neighbourhood  of  the  parotid  gland,  the  larynx,  and  pharynx, 
and  were  similar  in  every  respect  to  the  affection  of  the  jaw.  They 
were  described  as  starting  apparently  from  lymphatic  vessels  in 
these  parts.  Bollinger  discovered  the  fungus  in  a  case  of  so-called 
fibroid  of  the  second  stomach  of  a  cow,  a  spongy  growth  nearly  the 
size  of  the  fist ;  and  he  believed  that  in  another  case  the  disease 
manifested  itself  in  the  form  of  tubercular  ulceration  of  the 
intestines. 

Bollinger  submitted  the  fungus  to  Dr.  Harz,  a  botanist,  who 
described  the  fungi  as  mulberry-like  masses  from  *5  to  1  mm.  in 
diameter.  They  appeared  to  the  naked-eye  as  opaque,  white  grains,. 
and  when  calcined  were  difficult  to  recognise.  On  slight  pressure 
the  tufts  of  the  fungi  fell  apart  into  segments  of  unequal  size,  each 
of  which  appeared  to  correspond  to  an  individual  fungus.  The 
latter  was  described  as  beginning  at  the  pointed  end  of  the  wedge, 
with  a  somewhat  cone-shaped  basal  cell,  which,  in  the  absence  of  a 
mycelium,  perhaps  took  its  place,  and  bore  a  great  number  of  short 
linked  hyphse.  At  the  ends  of  the  hyphse  there  were  oval,  globular, 
or  elongated  club-shaped  bodies,  the  reproductive  cells  or  gonidia. 

Cultivation  experiments,  and  inoculation  of  the  tongue  of  a  calf 
with  liquid  containing  the  micro-organism,  failed.  Harz  proposed 


ACTINOMYCOSIS.  421 

to  call  the  fungus,  from  its  ray-like  appearance,  actinomyces  ;  but 
what  the  position  of  the  fungus  in  nature  might  be,  was  difficult  to 
determine.  It  did  not,  he  believed,  belong  to  the  yeast  fungi,  but  to 
the  mould  fungi,  and  might  be  compared  to  Botrytis,  Monosporium, 
arid  Polyacti.s. 

Bollinger  concluded  that  there  could  be  no  doubt  that  actino- 
mycosis  occupied  an  important  position  in  the  pathology  of  cattle 
diseases.  As  further  evidence  of  the  prevalence  of  the  affection,  he 
remarked  that  Zippelius  of  Obernburg  had  observed  in  the  course  of 
about  ten  years'  practice  not  less  than  254  cases  of  lymphoma,  in 
the  neighbourhood  of  the  larynx  and  pharynx,  besides  157  cases 
of  disease  of  the  jaw  ;  and  Bollinger  says  that  he  had  very  little 
doubt  that  the  greater  part  of  the  former,  and  very  likely  all  the 
cases  in  the  jaw,  were  due  to  the  fungus  which  he  had  discovered. 
In  certain  parts  of  Eranconia,  according  to  a  communication  received 
from  Professor  Frank,  these  tumours  of  the  throat  were  extremely 
abundant  in  cattle. 

Bellinger's  researches  were  followed  by  those  of  Siedamgrotzky, 
and  later  by  a  communication  from  Johne.  Johne  described  the 
various  forms  of  the  disease  which  had  up  to  that  date  been 
recognised,  including  a  description  of  actinomycosis  of  the  bones 
of  the  jaws,  of  the  fauces,  of  the  larynx,  of  the  oesophagus,  of  the 
stomach  and  intestinal  canal,  and  of  the  udder.  He  carried  out  a 
series  of  experiments,  by  which  it  was  clearly  established  that  the 
disease  could  be  communicated  from  cattle  to  cattle.  Previously 
Bollinger,  Harz,  Perroncito,  Ponfick,  Siedamgrotzky,  and  Johne  had 
failed,  but  subsequently  by  employing  fresh  material  from  the 
living  animal,  both  Johne  and  Ponfick  succeeded. 

Siedamgrotzky  not  only  confirmed  Bellinger's  researches,  but  he 
described  the  presence  of  the  fungus  in  so-called  "  multiple  sarcomas '' 
of  the  mucous  membrane  of  the  O3sophagus.  Rabe  described  the 
presence  of  the  fungus  in  tumours  known  as  Winddorn,  and  pointed 
out  that,  in  at  least  one  case,  he  considered  that  the  disease  had 
been  carried  by  the  lymphatics.  There  were  eleven  subcutaneous 
tumours  in  a  row  on  the  face,  which  were  connected  by  swollen, 
rope-like,  lymphatic  vessels.  They  appeared  to  be  secondary  to  a 
growth  on  the  nostril,  the  size  of  a  hen's  egg. 

Perroncito  described  a  case  of  "  sarcoma  "  of  the  intestines  and 
stomach,  which  proved  to  be  actinomycosis. 

Many  additional  communications  were  made  on  the  subject  of 
this  disease.  Ponfick  produced  it  in  the  lungs  by  intravenous 
injection,  and  subsequently  three  cases  occurring  naturally  in  the 


422  INFECTIVE    DISEASES. 

practice  of  veterinary  surgeons  were  published.  They  not  only 
deserve  especial  mention,  but  as  this  form  of  the  disease  appears  to 
be  so  seldom  recognised,  they  will  be  given  in  detail. 

Plug  described  a  case  in  the  lungs.  The  cow  had  been  out  of 
health  for  four  weeks,  did  not  eat,  and  had  a  cough,  and  two  days 
previous  to  the  visit  had  become  rapidly  worse.  Schmidt  found 
dyspnoea  with  abdominal  respiration ;  the  nostrils  were  dilated,  the 
head  protruded,  and  the  mouth  kept  open.  There  was  dulness  on 
percussion,  and  crepitation.  The  animal  was  killed,  and  the  lungs, 
which  alone  were  diseased,  were  sent  to  Plug.  The  pleura  on  exami- 
nation was  normal,  but  beneath  it  were  numbers  of  miliary 
tubercles,  many  equal  in  size  to  a  pin's  head.  On  section  the  lung 
had  a  granular  appearance  from  the  presence  of  countless  numbers 
of  minute  deposits,  which  all  had  the  appearance  of  grey  tubercles  ; 
in  none  was  there  any  central  softening.  They  were  present  in 
enormous  numbers  around  the  bronchi,  and  in  the  vessels  of  the 
interlobular  tissue.  Microscopical  examination  showed,  in  the  middle 
of  most  of  these  nodules,  the  presence  of  greenish-yellow,  radiating 
bodies,  which  under  a  high  power  appeared  to  be  undoubtedly 
actinomycotic  granules.  In  many  there  were  only  rudimentary  fungi 
consisting  of  four  or  five  clubs  ;  there  was  only  one  rosette  in  each 
tubercle.  The  fungus  was  surrounded  by  round  cells  and  fibrous 
tissue.  Larger  nodules  resulted  from  the  agglomeration  of  several 
tubercles,  or  from  diffuse  infiltration  of  round  cells  in  the  neighbour- 
hood of  a  tubercle. 

Hirik  met  with  a  somewhat  similar  case.  A  ten-j^ear-old  cow 
was  slaughtered,  and  in  the  middle  lobe  of  the  right  lung  there  were 
yellowish  nodules  about  the  size  of  a  pea,  scattered  over  an  area 
the  size  of  the  palm  of  the  hand.  These  nodules  were  not  at 
first  sight  distinguishable  from  ordinary  tubercles,  but  on  closer 
inspection  they  appeared  to  be  somewhat  different,  and  could  be 
easily  shelled  out  from  the  thickened  lung  tissue.  On  making  a 
section,  pus  welled  up  at  several  points,  and  contained  yellowish, 
calcareous  particles.  These  particles,  on  microscopical  examination, 
were  found  to  be  strongly  calcified  tufts  of  the  actinomyces  embedded 
in  granulation  cells.  Addition  of  hydrochloric  acid  dissolved  the 
calcareous  matter,  but  had  no  action  on  the  fungus. 

Pusch  described  a  third  case.  The  lungs  of  a  cow,  which  had 
been  killed  on  suspicion  of  having  pleuro-pneumonia,  were  sent  for 
examination.  The  front  lobe  of  the  left  lung  was  collapsed  and 
firm,  the  pleura  was  thickened  and  opaque  ;  the  larger  bronchi  were 
enlarged,  filled  with  pus,  and  their  walls  thickened.  In  the  posterior 


ACTINOMYCOSIS.  423 

lobe  of  the  left  lung  there  was  a  cavity  the  size  of  the  fist,  which 
had  been  opened,  and  the  contents  had,  for  the  most  part,  escaped  mf 
what  remained  was  a  greyish,  purulent  liquid,  full  of  yellowish 
bodies.  By  the  side  of  this  cavity  there  was  another  collection  of 
pus,  the  size  of  a  walnut.  In  the  lower  part  of  the  second  lobe  of 
the  right  lung  there  was  a  firm,  grey  tumour,  the  size  of  a  hen'* 
egg,  over  which  the  pleura  was  much  thickened.  On  section  thi& 
was  cavernous,  with  similar  purulent  contents,  and  yellow  grains. 
These  grains  under  the  microscope  proved  to  be  ray-fungi.  The  wall 
of  the  cavity  consisted  of  dense  connective  tissue  lined  with  a  soft 
granulation  tissue,  bathed  in  pus.  There  was  no  disease  of  any 
other  parts  in  'this  casa,  so  that  it  corresponded  in  this  respect  with 
the  two  previous  ones.  Pusch  adds  that  it  was  difficult  to  determine 
whether  the  organism  had  gained  access  to  the  lungs  by  the  blood- 
vessels, or  by  the  inspired  air.  In  his  case  he  inclined  to  the  latter 
view,  and  concludes  by  saying  that  the  organism  is  probably  very 
common  and  attached  to  the  most  varied  objects,  from  which  it 
may  be  conveyed  by  the  air. 

Pusch  refers  in  the  same  paper  to  an  interesting  case  which 
occurred  in  the  practice  of  Eggeling.  The  latter  had  under  his  care 
a  cow  with  extensive  paralysis.  The  spinal  cord  was  compressed 
by  a  compact  swelling  in  the  neck,  consisting  of  the  nodules  of 
actinomycosis.  There  were  no  manifestations  of  disease  in  any  other 
part  of  the  body. 

Prevalence  of  the  Disease. — The  author  found  that  the  disease 
was  not  generally  recognised  as  a  common  affection  of  cattle 
in  this  country,  in  spite  of  the  interest  excited  by  the  work  of 
Fleming,  to  wiiom  is  due  the  credit  of  first  recognising  a  case  in 
England.  In  1887  there  was  a  disease  prevailing  in  Norfolk,  and 
in  the  following  year  outbreaks  were  investigated  by  the  author 
in  Essex,  Hertfordshire,  Cambridgeshire,  and  Middlesex.  In  the 
Norfolk  outbreak  the  author  found  on  one  farm  8  per  cent,  of  the 
beasts  affected  with  the  so-called  "  wens "  or  "  sitfasts,"  which 
proved  on  microscopical  examination  to  be  cases  of  actinomycosis. 
These  growths  had  previously  been  described  in  veterinary  text-books 
as  the  result  of  strumous  or  scrofulous  inflammation ;  but  in  all 
the  specimens  of  wens  received  from  this  country  and  the  colonies, 
the  author  has  been  able  to  demonstrate  the  presence  of  the  ray- 
fungus. 

A  case  of  pulmonary  actinomycosis,  with  grape-like  growths  on 
the  pleura,  indicated  that  wens  were  not  the  only  manifestation  of 
this  disease,  which  had  teen  lost  sight  of  under  the  designation  of 


424 


INFECTIVE   DISEASES. 


tuberculosis.      Many  other  cases  were  examined,   and  the  disease 
was  shown  to  be  prevalent  in  this  country. 


FIG.  178. — From  a  photograph  of  a  Norfolk  steer.  There  is  a  growth  about  the 
size  of  an  orange  in  front  of  the  throat,  an  example  of  a  so-called  "  scrofulous  " 
or  "strumous"  tumour.  This  growth  was  associated  with  a  large  polypoid 
growth  in  the  pharynx  which,  by  interference  with  deglutition,  produced 
emaciation  (Fig.  180). 

In    Australia    actinomycosis    commonly  occurs  in    the    form   of 
tumours    of  the  upper   and  lower  jaw,    which  were  attributed  to 

"  cancer  "  or  to  "  scrofulous 
inflammation."  The  disease 
is  still  commonly  known  in 
Australia  as  "  cancer  "  and 
"  lumpy  jaw." 

Reports  of  the  prevalence 
of  actinomycosis  in  the  United 
States  have  been  published, 
by  the  Board  of  Live  Stock 
Commissioners  for  the  State 
of  Illinois.  In  their  Report 
for  1890  several  interesting 
communications  were  pub- 
lished. Mr.  Casewell,  State 
Veterinarian,  investigated  an 
outbreak  of  this  disease,  known  also  in  America  as  "  lumpy  jaw,' 


FIG.   179.— A  NORFOLK    HEIFER   WITH    A 
LARGE  '  WEN  "  IN  THE  PAROTID  REGION. 


ACTINOMYCOSIS. 


425 


-on  a  farm  in  Yates  City,  where  there  were  80  head  of  cattle,  and 
1C  were  found  to  be  suffering  from  actinomycosis.      Mr.  Casewell 


FIG.  180.— Photograph  of  a  steer  nearly  three  years  old,  but  about  the  size  of  a 
yearling.  The  emaciation  and  deplorable  aspect  recall  the  appearance  of 
"  a  piner  "or  "  waster  "  (tuberculosis). 

reported  that  the  disease  was  prevalent  in  nearly  every  county  in 
that  State,  and  that  in  his  opinion  it  was  spreading.  In  one 
instance  109  cases  were  slaughtered. 

Actinomycosis  in  Relation  to  Tuberculosis. — When  we  consider 
the  very  high  percentage  of  cases 
of  tuberculosis  which  has  been 
reported  in  some  localities,  the  im- 
portance of  differentiating  actino- 
mycosis from  tuberculosis  cannot 
be  over-estimated.  The  very 
great  contrast  in. the  appearance 
of  the  micro-organisms  in  the 
two  cases  renders  this  a  very 
easy  matter  for  the  pathologist. 
But  practical  veterinarians  and 
breeders  of  cattle  are  liable  to 
mistake  some .  manifestations  of 

actinomycosis  for  tuberculosis. 

T,     .         -     ,,  FIG.    181.— ACTIXOMYCOTIC    NODULES 

It    is    of    the    greatest    im-  FROM  THE  PLEURA. 

portance  to  bear   in  mind  that 

wens  or  clyers  are   really  not  tubercular,  but  actiiiomycotic ;  and 


426  INFECTIVE   DISEASES. 

that  a  condition  of  the  lungs  may  occur  as  the  result  of  actino- 
mycosis,  which  from  the  naked-eye  appearances  may  be  mistaken  for 
"grapes"  or  "  angleberries."  It  will  be  well  also  to  remember  in 
connection  with  the  above  remarks,  that  extreme  emaciation  may 
result  in  actinomycosis,  producing  a  condition  which,  without  a 
post-mortem  examination,  would  probably  be  attributed  to  tuber- 
culosis, the  animal  being  regarded  as  a  "  piner "  or  "  waster." 
If  these  possible  fallacies  are  taken  into  account,  the  excessive  per- 
centage of  tubercular  cases  so  commonly  reported  will  be  very 
considerably  reduced. 

There  is  no  evidence  to  show  that  the  flesh  of  animals  suffering 
from  actinomycotic  tumours  is  unfit  for  human  consumption.  In 
very  severe  cases  it  is  unwholesome,  but  there  is  no  evidence  that 
it  can  produce  actinomycosis  in  man. 

MANIFESTATIONS  OF  ACTINOMYCOSIS  IN  MAN. 

(I.)  Invasion  by  the  Mouth  and  Pharynx. — The  fungus  may  gain 
access  through  carious  teeth,  or  wounds  or  fistulse  of  the  jaw,  and 
very  possibly  by  inflammatory  processes  in  the  pharynx  and  tonsils. 

The  disease  attacks  the 'lower  jaw  most  frequently.  The  turnout- 
is  found  in  close  connection  with  the  bone,  or  in  the  sub-maxillary 
or  sub-mental  regions,  and  also  in  the  prse-tracheal  region.  It  occurs, 
though  rarely,  in  the  interior  of  the  bone. 

In  a  case  described  by  Israel,  which  occurred  in  a  woman  aged 
forty- six,  there  was  a  small  tumour  about  the  size  of  a  cherry 
attached  to  the  external  surface  of  the  lower  jaw,  with  an  opening 
through  which  a  probe  could  be  passed  into  the  bone.  The  tumour 
was  incised  and  scraped  away,  and  a  cavity  discovered  in  the  bone, 
admitting  a  small  sharp-spoon.  Later,  a  further  operation  was 
performed  :  the  periosteum  was  detached,  the  cavity  of  the  bone 
enlarged,  and  the  contents  scraped  out,  consisting  of  granulation 
tissue,  fragments  of  bone,  and  the  yellowish  fungi.  At  the  bottom 
of  the  cavity  the  fang  of  the  canine  tooth  was  found.  ISTo  return  of 
the  growth  occurred. 

The  first  cases  of  actinomycosis  which  were  observed  in  America 
were  connected  with  the  jaw.  In  1884  Dr.  Murphy  described  two 
cases  at  Chicago.  The  first  was  that  of  a  woman  aged  twenty-eight. 
Two  weeks  previously  she  had  suffered  from  severe  toothache,  with 
swelling  in  the  throat  and  great  pain  in  swallowing.  It  disappeared 
after  poulticing,  but  she  was  again  attacked  with  toothache,  and  a 
swelling  appeared  on  the  angle  of  the  jaw  on  the  left  side. 


ACTINOMYCOSIS.  427 

mouth  could  not  be  opened  without  difficulty ;  the  tonsil  was  much 
enlarged,  and  pus  was  set  free  on  incision.  She  still  suffered  with 
toothache,  and  a  small  swelling  now  occurred  on  the  left  side  of  the 
neck  below  the  jaw.  She  had  several  carious  teeth.  The  swelling, 
which  was  about  the  size  of  a  walnut,  was  punctured,  and  a  drainage 
tube  inserted  ;  a  creamy-looking  discharge  containing  yellow  granules 
continued  to  escape,  but  the  swelling  and  induration  increased.  A 
further  operation  was  decided  upon.  The  carious  tooth  was  removed, 
and  a  probe  passed  into  the  alveolus  showed  a  communication  with 
the  external  wound ;  the  angle  of  the  jaw  was  chiselled  aw~ay,  and 
the  alveolus  scraped  out.  lodoformed  gauze  was  applied,  and  the 
case  recovered. 

The  second  case  was  a  man  aged  eighteen,  who  had  also  suffered 
with  severe  toothache  and  swelling  at  the  angle  of  the  jaw.  On 
examination  a  carious  tooth  was  noticed.  The  swelling  was  well 
marked,  and  there  was  fluctuation  ;  it  was  as  large  as  a  pigeon's 
egg,  and  situated  below  the  jaw.  When  punctured,  thick  creamy 
pus  escaped  containing  the  fungi ;  the  sinus  was  scraped  out,  and  in 
ten  days  the  wound  was  healed.  Another  swelling  appeared,  and 
this  was  treated  as  before,  and  the  case  recovered. 

The  peculiar  feature  of  these  growths  is  their  apparent  migra- 
tion. Israel  states  that  in  one  case  a  tumour  occurred  on  the  alveolar 
process,  close  to  carious  teeth,  and  later  was  close  to  the  edge  of 
the  jaw  in  the  sub-maxillary  region.  From  thence  it  disappeared, 
and  a  large  swelling  formed  below  the  hyoid  bone,  and  after  this  had 
been  incised  and  had  healed,  an  abscess  formed  above  the  clavicle. 

Actinomycotic  tumours  in  this  region  would  sometimes  appear  to 
correspond  very  closely  with  wens  or  clyers  in  cattle  :  they  may  dis- 
charge through  the  skin,  and  the  opening  close,  or  a  fistula  result  ; 
but  they  differ,  from  their  tendency  to  form  burrowing  abscesses 
instead  of  recognisable  tumours.  In  this  respect  they  recall  chronic 
inflammation  rather  than  the  sarcoma-like  growths  in  cattle. 

Cases  in  which  the  upper  jaw  is  attacked  are  not  so  frequent  as 
those  in  the  lower  jaw.  The  progress  is  usually  described  as  slow, 
and  there  is  a  tendency  for  the  deep-seated  soft  parts  to  be  involved, 
while  in  the  lower  jaw  there  is  a  tendency  for  the  tumour  to  come 
to  the  surface.  There  may  be  burrowing  suppuration,  or  small 
tumours,  which,  after  a  time,  fluctuate  and  form  distinct  abscesses. 
These  may  involve  the  skin,  discharge  their  contents,  and  leave 
fistulous  openings. 

In  other  cases  the  disease  has  been  described  as  extending  from 
the  alveolar  process  to  the  temporal  bone,  or  the  base  of  the  skull, 


428  INFECTIVE    DISEASES. 

destroying  bones  and  even  reaching  the  brain ;  or  the  growth  may 
descend  by  the  spinal  column,  implicating  the  vertebrae,  and  travel- 
ling and  pointing  in  various  directions. 

(II.)  Invasion  by  the  Respiratory  Tract. — In  one  recorded  case 
the  disease  existed  for  seven  years,  was  localised  to  the  bronchi 
(Bronchitis  actinomycotica),  and  did  not  extend  into  the  lungs. 
The  sputum  was  examined,  and  contained  the  characteristic  fungus. 

If  the  micro-organisms  are  inhaled  they  pass  into  the  bronchioles 
and  alveoli,  and  produce  proliferation  of  round  cells,  which  undergo 
fatty  degeneration.  The  resulting  patches  of  peri-bronchitis  or 
pneumonia  become  yellowish-white ;  suppuration  and  haemorrhage 
from  the  capillaries  follow,  and  small  cavities  result,  containing  pus 
cells,  fat  granules,  blood,  and  the  fungi.  In  the  neighbourhood  of 
the  new  growth  there  is  compression  of  the  alveoli,  and  ultimately  the 
formation  of  a  dense  stratum  of  connective  tissue,  separated  from  the 
cavities  by  a  lining  of  granulation  tissue  containing  the  character- 
istic fungus.  The  symptoms  are  usually  obscure ;  but  the  sputum 
may  contain  the  fungi,  which  are  often  visible  to  the  naked  eye. 
The  apices  of  the  lungs  are  not,  as  a  rule,  affected.  There  is  con- 
siderable clinical  resemblance  to  chronic  phthisis  :  cough,  night- 
sweats,  pallor,  shortness  of  breath,  and  haemoptysis  are  symptoms 
common  to  both.  Light  may  be  thrown  upon  the  case  by  the  examina- 
tion of  the  sputum.  The  presence  of  the  actinomyces  will  be  positive 
evidence  as  to  the  nature  of  the  disease.  The  existence  of  these 
symptoms,  with  absence  of  tubercle  bacilli,  would  lead  to  the 
suspicion  of  actinomycosis,  even  failing  the  discovery  of  the  fungus 
in  the  sputum. 

In  the  second  stage  the  symptoms  are  more  characteristic.  The 
diseas3  spreads  to  neighbouring  parts,  and  pleurisy  commonly  super- 
venes. This  extension  may  involve  the  peri-pleural  tissues.  Thus 
the  disease  may  follow  the  prse-vertebral  tissues,  descend  behind  the 
insertion  of  the  diaphragm,  and  point  as  an  ordinary  psoas  or 
lumbar  abscess ;  it  may  perforate  the  diaphragm  and  reach  the 
abdominal  cavity.  Peritonitis  or  sub-phrenitic  abscess  may  then 
result.  In  some  cases  adhesions  have  formed,  and  the  disease  has 
extended  to  the  liver  or  spleen,  or  other  abdominal  organs.  The 
disease  may  also  extend  forwards  in  the  direction  of  the  anterior 
mediastinum  and  the  pericardium. 

The  primary  affection  of  the  lung  becomes  of  secondary  import- 
ance. Grave  symptoms  occur,  hectic  fever,  night-sweats,  rigors, 
and  marked  pallor.  In  the  third  stage,  the  disease  comes  to  the 
surface,  either  over  the  chest,  or  in  the  neighbourhood  of  the  dorsal 


ACT1NOMYCOSIS.  429 

or  lumbar  vertebrae  ;  a  swelling  appears  of  a  livid  colour,  and  if 
punctured  no  fluid  escapes,  but  if  allowed  to  make  its  own  way  to  the 
surface,  the  skin  gives  way,  a  muco-purulent  discharge  mixed  with 
pieces  of  the  growth  escapes,  and  the  fungi  can  readily  be  recognised. 

(III.)  Invasion  of  the  Digestive  Tract. — In  a  case  under  Chiari, 
death,  with  general  marasmus,  took  place  at  the  age  of  thirty-four, 
after  two  years'  illness.  The  mucous  membrane  of  the  intestines  was 
almost  completely  covered  with  whitish  patches,  raised  in  the  centre, 
and  covered  with  yellow  and  brown  granules  closely  adherent  ta 
the  adjacent  tissues.  The  teeth  were  carious. 

Small  nodules  about  the  size  of  a  pea  may  be  found  in  the  sub- 
mucous  tissue,  and  in  the  mucous  membrane  itself.  They  soften 
and  form  ulcers  with  undermined  edges,  the  base  reaching  the 
muscular  layer.  They  may  undergo  cicatrisation,  but  generally 
the  disease  extends  through  the  peritoneum  to  the  abdominal  cavity, 
and  perforates  the  bladder  or  the  intestines,  or  makes  its  way  through 
the  abdominal  wall.  Symptoms  are  either  absent  or  not  character- 
istic. The  fungus  may  sometimes  be  found  in  the  evacuations,  or 
by  exploratory  puncture. 

(IV.)  Undetermined. — In  addition  there  are  a  number  of  recorded 
cases  presenting  very  varied  symptoms  and  anatomical  relations,  in 
which  it  has  not  been  possible  to  satisfactorily  determine  the  path 
of  infection.  Delepine  has  described  a  most  interesting  case  of  an 
actinomycotic  tumour  of  the  brain. 

MANIFESTATIONS  OF  ACTINOMYCOSIS  IN  CATTLE. 

(I.)  In  the  Digestive  system  we  find  the  disease  attacking : — 

(a)  The   lips,  gums,    buccal  mucous  membrane   and  palate,    and 
appearing  as   nodules,  wart-like  growths,  or  ulcers.     The  nodules 
and  ulceration  of  the  palate  were  well  shown  in  a  specimen  sent  to 
the  author  for  examination,  under  suspicion  of  being  the  result  of 
severe  foot  and  mouth  disease. 

(b)  The  upper  and  lower  jaw,  where   it  probably  originates  in 
carious  teeth,  and  extending  and  invading  the  neighbouring  cavities 
and  sinuses  destroys   the  tissues  with  which  it  comes  in   contact, 
expanding   the   bones   into   thin    plates   or  reducing   them   to  the 
appearance  of  pumice-stone. 

(c)  The  tongue,  where  we  see  it  most  commonly  in  the  form  of 
nodules  or  wart -like  patches  under  the  mucous  membrane,  with  a 
special  tendency  to  ulcerate,  through  the  irritation  of  the  teeth.    These 
nodules  may  extend  into  the  deep  muscles,  and  often  collect  in  rows 


430 


INFECTIVE    DISEASES. 


more  or  less  parallel  to  the  superficial  muscular  fibres.  Complete 
tranverse  sections  of  the  tongue,  double-stained,  readily  show  this 
arrangement,  even  to  the  naked  eye.  Induration  of  the  tongue 
results  from  secondary  interstitial  glossitis.  The  author  has  seen, 
in  one  case  only,  a  tumour  embedded  in  the  substance  of  the  tongue 
about  the  size  of  a  small  Tangierine  orange,  and  more  or  less  isolated 
from  any  surrounding  growth. 

(d)  The  pharynx,  where  the  disease  may \  occur  in  the  form  of 
polypoid  growths  producing  asphyxia. 


FIG.  182. — A  NORFOLK  STEER  WITH  EXTENSIVE  ACTINOMYCOTIC  ULCER ATION  OF 

THE    SKIN  OF  THE  FLANK. 


(II.)  In  the  Respiratory  system  we  may  meet  with  the  disease  in  : — 

(a)  The  nasal  cavities,  originating  primarily  there  or  resulting 
from  extension  of  a  growth  from  the  lips,  or  the  pharynx. 

(6)  The  larynx  and  trachea,  generally  in  the  form  of  polypoid 
growths,  sessile  or  pedunculated,  which  arise  primarily  or  occur 
secondarily,  by  extension  from  the  tissues  in  the  neighbourhood. 

(c)  The  lungs,  where  the  differentiation  of  the  disease  is  most 
important,  as  neoplasms  in  the  lungs,  especially  in  the  early 
stages,  and  nodular  growths  on  the  pleura,  may  be  mistaken  for 
tuberculosis. 

The  disease  is  very  rarely  found  in  connection  with  the 
Nervous  system  (III.),  but  probably  does  not  so  rarely  attack  the 
Reproductive  system  (IV.). 


ACTINOMYCOSIS.  431 

(V.)  The  skin  and  subcutaneous  tissues  are  a  favourite  seat  of  this 
disease,  producing  the  so-called  wens  or  clyers  so  commonly  seen  in  the 
fen  country.  A  wen  is  first  recognised  as  a  small  tumour,  the  size 
of  a  marble  or  walnut,  which  increases  in  size  sometimes  with  great 
rapidity,  and  breaks  down  and  discharges  its  muco-purulent  contents 
through  the  inflamed  and  ulcerated  skin  ;  or  it  may  go  on  increasing, 
and  form  a  large  compact  growth,  the  size  of  a  child's  head.  These 
growths  when  excised,  hardened,  and  cut,  have  a  characteristic 
honeycombed  appearance,  produced  by  the  interlacing  bands  of 
fibrous  tissue,  which  form  a  spongy  structure,  from  the  interstices 
of  which  the  fungus  tufts  and  thick  yellowish  pus  have  for  the 
most  part  dropped  out. 

Actinomyces  Hominis.— Careful  examination  of  pus  from 
a,  case  of  actinomycosis  in  man  will  reveal  to  the  naked  eye  little 
yellowish-white  or  yellow  bodies,  which  a  casual  observer  might 
mistake  for  grains  of  iodoform.  On  collecting  some  of  the  discharge 
in  a  test-tube,  and  holding  it  between  the  light  and  the  eye,  the  tufts 
of  fungi  appeared  as  brownish  or  greenish-brown  grains,  embedded 
in  a  muco-purulent  matrix. 

On  spreading  some  of  the  discharge  on  a  glass  slip,  the  largest 
tufts  of  the  fungus  are  found  to  be  about  the  size  of  a  pin's  head. 
They  have  a  distinctly  sulphur- yellow  colour  by  reflected  light,  but 
appear  of  a  yellowish  or  greenish- brown  tint  by  transmitted  light. 
With  a  sewing  needle,  or  a  platinum  wire  flattened  at  the  end  into 
a  miniature  spatula,  the  grains  can  be  readily  picked  out  of  the 
discharge,  or  taken  off  the  dressing,  transferred  to  a  clean  slide,  and 
gently  covered  with  a  cover-glass.  Examined  with  an  inch  objective, 
they  have  the  appearance  of  more  or  less  spheroidal  masses  of  a 
pale  greenish -yellow  colour.  On  removing  the  preparation  from  the 
microscope,  and  gently  pressing  down  the  cover-glass  with  the  finger, 
the  grains  flatten  out  like  specks  of  tallow ;  and  on  again  examining 
with  the  same  power  they  are  found  to  have  fallen  apart  into  a 
number  of  irregular  and  sometimes  wedge-shaped  fragments  of  a 
faintly  brown  colour,  affording  a  characteristic  appearance.  By 
preparing  another  specimen,  and  covering  it  with  a  cover-glass 
without  completely  flattening  out  the  grains,  the  spherical,  oblong 
and  reniform  masses  of  which  the  tufts  are  composed  can  be 
recognised  with  a  J-  in.  objective  as  rosettes  of  clubs.  By  examining 
the  peripheral  part  of  a  rosette  with  a  Yj-in.,  and  especially  after 
pressing  the  grains  into  a  thin  layer,  with  or  without  the  addition 
of  a  drop  of  glycerine,  the  characteristic  clubs  are  most  readily 
demonstrated,  and  the  most  varied  shapes  observed  by  carefully 


432  INFECTIVE   DISEASES. 

examining  the  form  of  the  individual  elements.  As  in  the  bovine 
,  fungus,  every  variation  in  form  is  found,  from  single  clubs  to  clubs 
with  lateral  offshoots,  clubs  bifid  at  the  extremity,  palmate  or 
fan-shaped  groups,  and  banana-like  bunches.  In  many  cases  the 
clubs  are  divided  by  transverse  fission  into  two,  three,  or  more 
segments.  As  a  rule,  the  clubs  are  irregular  in  shape,  and  of  about 
equal  size,  while  a  few  are  conspicuous  by  their  length.  In  other 
parts  of  the  preparation  the  clubs  are  replaced  by  long  slender 
forms,  which  are  sometimes  transversely  divided  into  a  number  of 
short  links.  With  suitable  illumination  many  clubs  are  seen  to 
taper  off  into  slender  filaments.  In  addition  there  are  free  filaments, 
which  are  twisted,  branched,  and  sometimes  distinctly  spirilliform. 
Many  of  the  clubs  are,  composed  of  layers  differing  in  their  refractive 
power,  and  many  have  the  appearance  of  a  cejntrajjchannel.  There 
are  also  in  the  preparation  small,  highly  refractive  bodies,  fat 
granules,  granular  detritus,  round  cells,  pus  cells,  and  sometimes 
blood  corpuscles. 

The  grains  differ,  as  a  rule,  from  those  from  a  bovine  source,  in 
the  absence  of  that  sensation  of  grittiness  so  often  transmitted  to  the 
finger  when  pressing  the  cover-glass  upon  them,  and  in  the  slightly 
greater  tendency  of  the  tufts  to  retain  their  compact  form.  By 
teasing  the  grains  in  a  drop  of  water  on  a  slide,  and  examining 
the  preparation  with  a  -g-  or  a  ~%  objective,  the  explanation 
of  the  latter  is  forthcoming ;  for  by  this  process  the  clubs  are 
gradually  washed  away,  and  a  central  core  remains,  which  is  com?- 
posed  entirely  of  a  dense  network  of  filaments.  This  can  readily  be 
observed  by  using  a  small  diaphragm,  and  it  will  be  found  that 
the  rosettes  of  clubs  are  now  replaced  by  tangled  masses,  having  some 
resemblance  to  miniature  tufts  of  cotton-wool.  These  filaments 
constitute  the  delicate  network  which  is  seen  in  sections  stained 
by  the  method  of  Gram.  This  can  be  readily  verified  by  making 
a  cover-glass  preparation  of  the  grains,  and  staining  by  that  method. 
The  characters'  of  the  fungus  can  readily  be  studied  by  proper 
illumination,  without  staining.  The  clubs  have  a  faintly  greenish 
tint,  and  in  form  and  arrangement  are  quite  characteristic  and  easily 
recognisable.  Permanent  preparations  may  be  made  by  mounting 
the  fungus  in  glycerine. 

DESCRIPTION  OF  STAINED  SPECIMENS. 

The  fungus  may  be  stained  in  alcoholic  solution  of  eosin  in  the 
manner  to  be  described  for  the  bovine  organism,  or  in  orarige-rubinr 


DESCRIPTION   OF    PLATES   XV.    AND    XVI. 
Actinomyces. 

PLATE   XV. 

FIG.  1. — From  a  preparation  of  the  grains  from  an  actinomycotic  abscess  in 
a  boy ;  examined  in  glycerine.  The  drawing  has  been  made  of  a  com- 
plete rosette  examined  by  focussing  successively  the  central  and  peripheral 
portions.  Towards  the  centre  the  extremities  of  the  clubs  are  alone 
visible ;  they  vary  in  size,  and  if  pressed  upon  by  the  cover-glass  give  the 
appearance  of  an  irregular  mosaic.  Towards  the  periphery  the  clubs  are 
seen  in  profile,  and  their  characteristic  form  recognised.  At  one  part 
there  are  several  elongated  elements,  composed  of  separate  links,  x  1200. 
FIG.  2. — Different  forms  of  clubs  from  preparations  in  which  the  rosettes  have 
been  flattened  out  by  gentle  pressure  on  the  cover-glass,  x  2500. 

(a)  Single  club.  (#)  Bifid  club.  (0)  Club  giving  rise  to  four 
secondary  clubs.  (cT)  Four  clubs  connected  together,  recalling 
the  form  of  a  bunch  of  bananas,  (e)  Mature  club  with  a  lateral 
bud.  (/)  Apparently  a  further  development  of  the  condition 
represented  at  (e).  (^)  Club  with  a  lateral  bud  and  transverse 
segmentation.  (Ji)  Single  club  with  double  tranverse  segmenta- 
tion, (i)  Club  with  oblique  segmentation,  (j)  Collection  of 
four  clubs,  one  with  lateral  gemmation,  another  with  oblique 
segmentation.  (&)  Club  with  lateral  buds  on  both  sides,  and 
cut  off  square  at  the  extremity.  (Z)  Club  with  a  daughter  club 
which  bears  at  its  extremity  two  still  smaller  clubs,  (nt)  Club 
divided  by  transverse  segmentation  into  four  distinct  elements. 
(#)  Elongated  club  composed  of  several  distinct  elements.  (0)  and 
(jy~)  Clubs  with  terminal  gemmation.  (^)  Palmate  group  of  clubs. 
(r)  Trilobed  club,  (s)  Club  with  apparently  a  central  channel. 
(£)  Filament  bearing  terminally  a  highly  refractiveoval  body. 

PLATE    XVI. 

FIG.  1. — From  a  section  of  a  portion  of  the  growth  removed  from  a  boy 
during  life.  The  tissue  was  hardened  in  alcohol,  and  cut  in  celloidin. 
The  section  was  stained  by  Gram's  method  and  with  orange-rubin.  x  50. 

FIG.  2. — From  the  same  section.  A  mass  of  extremely  fine  filaments  occupies 
the  central  part  of  the  rosette.  Many  of  the  filaments  have  a  terminal 
enlargement.  The  marginal  part  shows  a  palisade  of  clubs  stained  by  the 
orange-rubin.  x  500. 

FIGS.  3  and  4. — From  cover-glass  preparations  of  the  fungus  teased  out  of  the 
new  growths  produced  by  inoculation  of  a  calf  with  pus  from  a  boy 
suffering  from  pulmonary  actinomycosis.  Stained  by  Gram's  method  and 
orange-rubin.  The  threads  are  stained  blue  and  the  clubs  crimson  (a) 
In  the  younger  clubs  the  thread  can  be  traced  into  the  interior  of  the 
club  (#).  In  some  of  the  older  clubs  the  central  portion  takes  a  yellowish 
stain,  and  in  others  the  protoplasm  is  not  continued  as  a  thread,  but  is 
collected  into  a  spherical  or  ovoid  or  pear-shaped  mass.  In  others,  again 
irregular  grains  stained  blue  are  scattered  throughout  the  central  portion 
(Fig.  4).  x  1200. 

FIG.  5.  From  a  pure-culture  on  glycerine-agar.  («)  branching  filaments,  (J)  a 
mass  of  entangled  filaments.  Gram's  method,  x  1200. 

FIG.  6. — From  a  similar  but  older  cultivation,  (a)  a  filament  with  spores 
(J)  chains  of  spores  simulating  streptococci.  Gram's  method,  x  1200. 


ACTINOMYCOSIS.  433- 

and  in  either  case  mounted  in  glycerine.  But  although  the  fungus 
can  be  detected  without  any  staining  process,  there  may  sometimes 
be  doubtful  appearances,  and  then  cover-glass  preparations  should  be 
made  and  stained  by  the  method  of  Gram  with  eosin.  The  filaments 
can  be  readily  recognised,  and  this  is  of  great  value,  as  it  forms 
an  additional  means  for  the  diagnosis  of  the  disease.  In  combination 
with  orange-rubin  we  have  a  test  that  is  as  characteristic  and  useful 
as  staining  for  tubercle  bacilli.  The  discharge,  scraping  from  a 
growth,  sputum,  or  the  isolated  fungus  is  squeezed  between  two- 
cover-glasses,  which  are  then  slid  apart ;  they  are  allowed  to  dry, 
passed  through  the  flame  in  the  ordinary  manner,  and  then  stained. 
The  cover-glasses  can  be  cleared  in  clove-oil,  the  excess  of  clove-oil 
being  removed  by  gentle  pressure  between  pieces  of  blotting-paper,, 
and  then  the  preparation  can  be  mounted  in  balsam  and  rendered  per- 
manent. On  examination  of  these  specimens  the  masses  of  filaments 
will  be  found  to  be  stained  blue,  and  the  tissue  elements  pink.  These 
filaments  vary  very  much  in  extent  and  character  in  different  pre- 
parations. In  some  cases  there  are  masses  of  short  threads,  which 
are  either  straight,  sinuous,  or  twisted,  and  branched.  In  other 
parts  the  field  is  occupied  by  very  short,  straight,  or  curved  and 
sometimes  spiral  fragments ;  in  others,  again,  there  are  comparatively 
long  strands.  On  examination  with  a  high  power,  and  with  careful 
illumination,  some  filaments  will  be  observed  to  be  moniliform, 
while  others  are  provided  with  a  terminal  oval  body.  There  are 
also  free  spherical,  and  oval,  bodies  stained  blue,  which  represent  the 
spores  of  the  organism.  When  orange-rubin  is  used  instead  of 
eosin,  the  clubs  will  be  stained  and  easily  recognised.  This  method 
enables  one  to  determine  the  exact  relation  of  the  threads  to  the- 
club-shaped  bodies ;  and  this  is  an  interesting  point,  as  it  has  been 
suggested  that  the  threads  are  not  connected  with  the  clubs,  but  are 
merely  an  adventitious  micro-organism  growing  in  the  track  of  the 
ray-fungus.  The  threads  are  stained  blue  and  the  clubs  crimson. 
In  the  younger  clubs  the  protoplasm  of  the  thread  can  be  traced  inta 
the  interior  of  the  club.  In  some  of  the  older  clubs  the  central 
portion  takes  a  yellowish  stain,  and  in  others  the  protoplasm  is  not 
continued  as  a  thread,  but  is  collected  into  a  spherical,  ovoid,  or 
pear-shaped  mass.  In  others  again,  irregular  grains,  stained  blue,  are 
scattered  throughout  the  central  portion.  The  sheath  of  the  thread 
is  stained  pink ;  and  the  protoplasm,  stained  blue,  fills  the  sheath,  or 
consists  of  small  spherical  or  irregular  grains,  giving  a  distinctly 
beaded  appearance. 

The  effect  of  various  reagents  should  be  tried  upon  the  isolated 

28 


434  INFECTIVE   DISEASES. 

grains.  The  grains  are  picked  out  of  the  pus  and  transferred  to 
watch-glasses  containing  strong  potash,  xylol,  and  benzol.  If 
returned  to  a  slide  and  covered  with  a  cover-glass,  the  clubs  are 
found  unaltered.  Water  or  weak  potash  washes  away  the  clubs, 
and  the  filaments  become  easily  distinguished  ;  ether  and  strong 
acids  have  no  effect  upon  them.  Corallin  soda,  Hanstein's  violet, 
and  iodine  zinc-chloride  fail  to  give  any  particular  reaction. 
Hoffman's  blue  stains  the  clubs,  but  without  bringing  out  any 
structural  details  which  could  not  be  observed  in  the  unstained 
specimens. 

Actinomyces  bovis. — The  fungus  in  cattle  may  in  the  same 
way  be  detected  with  the  naked  eye  in  the  muco-purulent  discharge, 
or  in  a  scraping  from  the  cut  surface  of  a  growth.  The  tufts  of  the 
fungus  vary  in  size  under  different  circumstances,  from  that  of  a  grain 
of  fine  sand  to  that  of  a  pin's  head.  If  the  pus  or  scraping  be  spread 
out  on  a  slide  and  examined  against  a  dark  background,  the  grains 
appear  to  be  white  or  yellowish-white  in  colour  ;  but  if  examined  by 
transmitted  light,  they  appear  distinctly  brownish.  On  pressing  the 
cover- glass  on  the  slide  the  grains  readily  flatten  out,  being  of  a 
soft,  tallowy  consistency ;  or  in  the  process  of  gently  pressing  the 
cover-glass  on  the  slide  with  slight  lateral  movement,  a  distinct 
gritty  sensation  is  transmitted  to  the  finger,  owing  to  the  presence 
of  calcareous  matter.  On  examination  with  a  low  power  the 
fungus  will  be  recognised  in  the  form  of  irregular  patches  scattered 
over  the  field,  which  might  readily  be  regarded  as  collections  of 
granular  debris  of  a  brownish  or  yellowish-brown  colour,  but  on 
careful  examination  they  are  observed  to  have  a  more  or  less 
characteristic  appearance.  On  examining  with  a  higher  power, 
spherical,  ovoid,  or  reniform  bodies  are  to  be  seen,  which  are 
either  typical  rosettes  of  clubs  or  granular  masses,  with  here 
and  there  a  club-shaped  body  at  the  periphery.  Pus  cells,  round 
cells,  fat  granules,  and  minute  spherical  bodies  may  also  be 
distinguished.  If  the  grains  consist  of  typical  rosettes,  and  be 
merely  covered  with  the  cover-glass,  and  examined  without  being 
flattened  out  between  the  cover-glass  and  the  slide,  they  will  recall 
to  mind,  on  focussing  alternately  the  centre  and  the  periphery, 
the  appearance  of  the  capituluin  of  a  composite  flower.  The  central 
portion  appears  to  consist  of  spherical  forms;  these  are  the  ex- 
tremities of  the  component  elements,  and  as  we  focus  the  edge  of 
the  rosette  these  elements  are  seen  laterally,  and  their  characteristic 
club-form  is  readily  distinguished.  The  central  portion  may  be 
flattened  against  the  cover-glass,  and  as  the  individual  clubs  vary 


DESCRIPTION    OF    PLATES    XVII.    AND    XVIII. 
Actinomycosis  Bovis. 

PLATE    XVII. 

Section  of  an  actinomycotic  tongue  stained  by  the  method  of 
Gram  and  with  eosin. 

FIG.  1. — This  illustrates  the  appearance  which  is  usually  seen  under  a  low 
power,  when  a  section  is  stained  by  Gram's  method  and  with  eosin.  The 
central  portion  of  a  mass  of  the  fungus  is  either  unstained  or  tinged  with 
eosin,  while  the  marginal  portion  is  stained  blue.  The  reverse  is  seen,  as  a 
rule,  in  sections  from  man  ;  although  under  a  low  power  the  general  appear- 
ance of  sections  from  these  two  sources  is  somewhat  similar,  x  50. 

FIG.  2.  —a,  b,  c>  d,  represent  the  earliest  recognisable  forms  of  the  ray  fungus 
in  the  interior  of  leucocytes.  In  e  the  club-forms  can  be  recognised.  In 
f  and  g  there  are  small  stellate  groups  of  clubs,  x  500. 

FIG.  3. — A  part  of  the  section  represented  in  Fig.  1,  under  a  high  power.  The 
marginal  line  of  blue  observed  under  a  low  power  is  now  recognised  as  the 
result  of  the  stain  being  limited  to  the  peripherally  arranged  clubs.  At 
(«)  part  of  a  rosette  has  undergone  calcification  ;  the  clubs  are  granular, 
and  have  not  retained  the  stain.  At  (J)  and  close  to  it  there  are  the 
remains  of  rosettes  in  which  the  process  of  calcification  is  almost  complete. 
x  500. 

PLATE    XVIII. 

The  figures  in  this  plate  are  taken  from  sections  of  a  case  of 
so-called  "  osteosarcoma,"  in  which  the  growth  of  the  fungus  was 
remarkably  luxuriant.  The  specimens  were  stained  by  Plants' 
method. 

FIG.  1.— Different  forms  of  clubs  in  different  specimens  :    x  1200. 
O)  Very  small  club-shaped  elements. 
(&)  A  club  with  transverse  segmentation, 
(c)  A  club  with  lateral  daughter  clubs. 

(d  and  <?)  Clubs  with  terminal  offshoots  resembling  teleutospores. 
(/)  A  club  with  developing  daughter  clubs  on  the  left,  and  on  the 

right  a  mature  secondary  club. 
(# )  A  segmental  club  with  lateral  offshoots. 
(1C)  Two  clubs  undergoing  calcification. 

FlG.  2.— A  very  remarkable  stellate  growth  comprised  of  nine  wedge-shaped 
collections  of  clubs  radiating  from  a  mass  of  finely  granular  material. 
x  500. 

FIG.  3.— A  rosette  undergoing  central  calcification,  and  consisting  in  part  of 
extremely  elongated  clubs  resembling  paraphyses.  Calcareous  matter  is 
also  being  deposited  in  the  club-shaped  structures,  x  500. 
FIG.  4. — Part  of  a  rosette  with  continuation  of  the  club-shaped  bodies  into 
transversely  segmented  branching  cells  apparently  representing  short 
hyphae.  x  500. 

FIG.  5. — A  rosette  from  another  section  in  which  similar  appearances  are 
observed  as  in  Fig.  4.  x  500. 


Plate  XV11 


T-igl 


* 


SIS   BOX"  IS 


Hnaml  flrecfc^io-.    I 


Plate  XVIII 


BOVIS 


.  t&> 


ACTINOMYCOSIS.  435 

considerably  in  size,  the  appearance  of  an  irregular  mosaic  Is  thus 
produced. 

By  pressing  upon  the  cover  we  break  up  the  rosette,  and  then 
the  clubs  are  recognised  either  singly  or  in  pairs,  or  attached  together 
in  the  form  of  wedge  or  fan-shaped  segments.  Calcareous  material, 
if  present,  may  readily  be  demonstrated  by  the  action  of  acids.  It 
will  be  found  that  on  the  addition  of  dilute  hydrochloric,  nitric  or 
acetic  acids,  the  calcareous  deposit  is  dissolved  while  the  clubs  are 
not  affected,  and  even  with  the  addition  of  the  strongest  acids  the 
only  re"'  it  will  be  to  dissolve  out  the  calcareous  matter  and  clarify 
the  tutcs  of  the  fungus,  the  form  of  the  clubs  being  still  recognisable. 
They  are  not  affected  by  ether  or  potash,  and  thus  the  effects  of 
chemical  reagents  clearly  distinguish  them  from  fat  crystals  or 
calcareous  particles.  By  breaking  up  the  growth  into  small  frag- 
ments, we  may  readily  study  the  shape  of  the  individual  club-like 
•elements.  By  using  high-power  objectives,  and  properly  arranging 
the  illumination,  various  forms  will  be  clearly  delineated.  In  some 
cases  the  club  will  be  found  to  be  bifid  at  the  extremity  ;  in  other 
cases  there  are  lateral  offshoots  or  daughter- clubs.  Here  and  there 
will  be  found  clubs  closely  pressed  together  like  a  bunch  of  bananas, 
and  in  other  cases  the  broken-off  pieces  have  a  palmate  form.  By 
teasing  out  the  grains  in  water,  and  pressing  them  apart  between 
the  slide  and  cover-glass,  we  find  that  the  central  portion  is  com- 
posed, as  a  rule,  of  a  structureless  core.  More  rarely  there  are  the 
delicate  filaments  which  are  found  in  cultures  and  in  the  fungus  from 
man. 

If  the  grains  are  mounted  in  glycerine,  the  appearance  of  the 
organism  in  the  fresh  state  may  be  preserved. 

The  granules  may  be  stained  by  picking  them  out  with  needles 
and  transferring  them  to  a  watch-glass  containing  alcohol,  to  which  a 
few  drops  of  concentrated  alcoholic  solution  of  eosin  have  been  added. 
They  remain  in  the  solution  until  distinctly  stained,  and  they  are 
then  placed  on  a  glass  slide  in  a  drop  of  glycerine. 

The  muco-pus  may  be  spread  out  into  as  thin  a  film  as  possible 
on  a  cover-glass,  allowed  to  dry,  fixed  by  warming  slightly  over 
the  flame,  and  stained  by  the  method  of  Plant  or  of  Gram.  The 
characters  of  the  fungus  can  be  so  readily  recognised  in  the  perfectly 
fresh  state,  that  methods  of  staining  are  of  secondary  importance 
in  diagnosis,  though  there  are  certain  minute  points  which  can  only 
be  satisfactorily  determined  by  means  of  suitable  dyes. 


436  INFECTIVE    DISEASES. 

.CULTIVATION  OF  ACTINOMYCES. 

Bostrb'm  cultivated  actinomyces  from  five  cases  in  animals,  and 
from  one  case  in  man.  In  all  cases  he  obtained  a  similar  result. 
The  fungi  were  isolated  from  pus  with  sterilised  needles,  and  placed 
in  liquefied  nutrient  gelatine  in  which  they  were  teased  out,  and  the 
gelatine  then  spread  on  glass  plates.  The  growth  is  stated  to  have 
become  visible  in  a  few  days.  The  fungi  were  isolated  from  the 
plates,  crushed  between  sterilised  glass  slides  arid  inoculated  on  the 
surface  of  nutrient  agar-agar  and  blood  serum.  In  this  way  pure 
cultivations  were  obtained.  Nutrient  gelatine  was  not  liquefied. 
The  cultures  in  blood  serum  and  agar-agar  grew  best  at  from  33° 
to  37°  C.  The  track  of  inoculation  gradually  spread  out  during 
the  first  two  days,  having  a  finely-granular,  whitish  appearance. 
During  the  next  few  days  small  yellowish-red  spots  appeared  in  the 
centre  of  the  inoculated  area,  while  the  edge  was  apparently  com- 
posed of  fine  processes.  The  yellowish  spots  continued  to  increase- 
for  about  seven  or  eight  days,  and  became  confluent,  and  the 
periphery  also  was  dotted  with  yellowish-red  points.  Finally,  there 
were  also  isolated  colonies  consisting  of  a  yellowish-red  centre  with 
a  greyish,  downy  periphery.  The  cultivated  fungus,  if  suitably 
stained,  corresponded  exactly  with  that  found  in  human  and 
animal  actmornycosis.  In  the  cultures  during  the  first  two  days 
threads  were  found,  with  true  branchings  ;  later  the  threads  were 
divided  into  shorter  pieces  or  rods,  and  when  the  yellowish  centres 
appeared  there  were  also  a  number  of  very  short  rods  and  cocci- 
like  forms.  Bostrom  also  described  attenuated  club-shaped  syell- 
ings  at  the  end  of  the  threads.  He  concluded  by  saying  that 
Actinomyces  is  not  one  of  the  mould  fungi,  the  central  threads  do 
not  therefore  constitute  a  mycelium ;  he  was  inclined  to  regard  it 
as  a  branched  Cladothrix,  and  cultivation  seemed  to  prove  this.  He 
suggested  that  it  might  be  the  Streptothrix  Forsteri  of  Colin.  Tii 
any  case  he  relegated  Actinomyces  to  the  fission  fungi  or  bacteria. 

In  1888  the  author  made  cultures  on  glycerine-agar  from  a  case  of 
human  actinomycosis  of  the  thoracic  wall.  An  abscess  was  opened,  the 
discharge  collected  in  sterilised  tubes,  and  cultivations  prepared  with 
as  little  delay  as  possible.  Some  of  the  discharge  was  spread  out  on 
a  sterilised  glass  slide,  and  the  grains  isolated  with  sterilised  needles 
and  quickly  transplanted  on  the  surface  of  the  nutrient  medium. 
The  tubes  were  placed  in  the  incubator  at  37°  C.,  and  the  result 
watched  from  day  to  day.  For  several  days  there  was  to  the 
naked  eye  no  promise  of  success;  but  gradually  the  grains  began 


* 

ACTINOMYCOSIS.  437 

to  change,  and  by  the  end  of  a  fortnight  there  was  an  appreciable 
increase  in  size.  Numerous  cover-glass  preparations  were  made 
from  what  was  originally  a  single  grain,  and  on  examination  by  the 
method  of  Gram  the  appearance  was  very  striking.  There  could  be 
no  doubt  as  to  the  increase  of  the  mycelial  structure.  The  dense 
masses  of  filaments  covered  almost  the  whole  area  of  the  preparation. 
In  parts  less  thickly  covered  there  were  very  numerous  oval 
bodies,  and  rod-like  segments  with  terminal  enlargements.  These 
" crocus"  forms  corresponded  with  the  appearances  previously  de- 
scribed as  met  with  in  the  interior  of  certain  clubs.  From  this  it 
would  appear  that  some  other  condition  is  necessary  for  the  develop- 
ment of  the  fully  formed  club,  which  is  the  result  of  the  sheath 
undergoing  some  change,  possibly  mucilaginous,  resulting  in  the 
formation  of  a  thick  investment  of  the  clubbed  mass  of  protoplasm 
at  the  end  of  the  thread. 

These  club-shaped  bodies  represent  organs  of  fructification,  rather 
than  the  results  of  degeneration  or  death.  The  difficulty  in  accept- 
ing the  view  of  their  being  entirely  lifeless  forms  lies  in  the  fact  that 
the  author  has  observed  daughter-clubs  growing  from  the  mature 
clubs ;  and,  further,  in  the  bovine  fungus  the  author  has  been 
able  to  trace  the  stages  in  the  development  of  a  single  club  to  a 
completely  formed  rosette. 

In  the  unstained  condition,  the  clubs  are  found,  on  the  whole,  to 
be  very  regular  in  their  form  and  arrangement,  and  by  certain 
staining  methods  they  can  be  shown  to  have  a  somewhat  complex 
structure.  If  we  take  all  the  characters  into  account,  and  par- 
ticularly the  minute  structure  and  the  relation  to  each  other  of  the 
threads  and  clubs,  we  are  justified  in  the  opinion  that  the  club  in 
the  early  stages  is  an  integral  part  of  the  living  fungus,  and  that 
these  characters  bring  the  fungus  into  relation  with  a  higher  group 
of  micro- fungi,  Basidiomycetes,  although  the  filaments,  regarded 
by  themselves,  correspond  with  the  characters  of  Streptothrix.  The 
life-history  of  the  micro-organism  may  be  summed  up  thus  : — 

The  spores  sprout  into  excessively  fine,  straight,  or  sinuous,  and 
sometimes  distinctly  spirilliform  threads,  which  branch  irregularly 
and  sometimes  dichotomously.  The  extremities  of  the  branches 
develop  the  club-shaped  bodies.  The  clubs  are  closely  packed  to- 
gether, so  that  a  more  or  less  globular  body  is  formed,  with  a  central 
core  composed  of  a  dense  mass  of  threads.  The  threads  can  be 
differentiated  by  the  method  of  Gram  into  an  external  sheath,  and 
protoplasmic  contents.  The  club-shaped  body  externally  appears  to 
be  mucilaginous,  while  internally  it  is  continuous  with  the  protoplasm 


I 

438  INFECTIVE    DISEASES. 

of  the  thread.  It  is  difficult  to  say  what  further  changes  occur  in 
the  club-shaped  bodies  ;  in  all  probability  they  represent  organs  of 
fructification.  If  so,  the  protoplasm  in  the  interior  of  the  club  may 
possibly  undergo  changes  leading  to  the  development  of  spores, 
which  are  ultimately  set  free ;  in  some  cases  the  terminal  segment 
of  a  club  is  separated  by  transverse  fission  in  the  form  of  a  globular 
body,  a  process  resembling  the  formation  of  spores  by  abjunction. 
In  others,  the  forms  sprouting  from  the  club  are  suggestive  of 
teleutospores.  There  are  occasionally  long,  slender  forms,  very 
different  from  the  ordinary  clubs  ;  they  possibly  represent  paraphyses 
or  abortive  elements.  In  whatever  way  they  may  be  formed,  there 
can  be  little  doubt  that  spores  are  set  free  in  the  vicinity  of 
a  rosette,  and  give  rise  to  fresh  individuals;  the  ultimate  result 
recalling,  as  has  been  suggested,  the  appearance  of  "  fairy  rings." 
There  can  be  little  doubt  that  spores  and  young  fungi  are  taken 
up  by  wandering  cells,  and  conveyed  to  a  distance  from  the  parent 
fungus,  and  thus  fresh  centres  of  growth  are  established. 

Appearances  of  cultures. — Bostrom,  Wolff,  Israel,  Paltauf,  and 
others  have  shown  that  actinomyces  can  be  cultivated  in  the 
ordinary  nutrient  media.  More  recently  the  author  has  carried  on 
a  series  of  cultivations  for  some  years  on  glycerine  agar,  gelatine 
and  milk,  broth,  bread-paste,  and  potato,  in  order  to  observe  the 
changes  which  take  place,  and  to  study  the  variations  which  he 
found  in  the  appearance  of  sub-cultures.  The  actinomyces  after  a 
few  days  on  glycerine  agar  at  the  temperature  of  the  blood  forms 
little,  white,  shining,  moist  colonies,  which  may  remain  stationary,  or 
increase  and  coalesce.  In  a  week  or  ten  days,  sometimes  earlier,  and 
sometimes  after  several  weeks,  the  cultures  turn  a  bright  yellow  colour, 
but  some  remain,  though  white ;  others,  again,  have  a  tinge  of  pink, 
and  others  are  yellowish-brown  (Plate  XIX).  After  a  time,  a  powdery 
efflorescence  makes  its  appearance  on  the  surface  of  the  culture, 
which  may  be  either  yellow  or  white  in  colour.  The  culture  may  go 
on  increasing,  spreading  over  the  surface  of  the  medium,  and  retain 
its  yellow  colour,  or  it  may  turn  black  in  parts  or  completely  so, 
while  the  agar  is  coloured  brownish-black.  Cultures  have  a  peculiar 
sour  smell;  the  variations  in  cultures  were  all  proved,  by  careful 
testing  by  sub-cultures,  to  be  due  to  the  growth  of  the  actinomyces 
under  varying  conditions  of  soil,  temperature,  and  the  supply  of  air. 
The  stage  of  efflorescence  corresponds  with  the  breaking  up  of  the 
filaments  into  masses  of  cocci,  and  chains  often  closely  resembling 
streptococci.  Gelatine  is  slowly  liquefied. 

Wolff   and   Israel    cultivated   actinomyces    on   raw   and   boiled 


DESCRIPTION    OF    PLATE    XIX. 
Pure-cultivations  of  Actinomyces. 

These  tubes  were  selected  from  a  great  number  of  cultivations 
in  which  there  were  different  appearances.  In  some  instances  the 
growths  had  a  faint  tinge  of  pink. 

FIG.  1.— Pure-cultivation  on  the  surface  of  potato,  showing  a  luxuriant 
sulphur-yellow  growth  entirely  composed  of  entangled  masses  of  fila- 
ments. After  three  months'  growth. 

FIG.  2. — Pure-culture  from  the  same  series,  on  glycerine-agar.  In  this  case 
the  culture  remained  perfectly  white.  The  jelly  was  coloured  reddish- 
brown.  After  fifteen  months'  growth. 

FIG.  3. — Pure-culture  on  glycerine-agar  in  which  the  growth  was  dark- 
brown,  in  parts  black,  and  the  jelly  stained  dark-brown.  After  nearly 
two  years'  growth. 


PlateHX. 


Figl.  Fig  2. 

PURE- CULTIVATIONS 

OF 
ACT  INOMYCES. 


Vbifgnl  Bnolcs.Day  £Son.,IitK. 


ACTINOMYCOSIS.  439 

eggs,  and  succeeded,  by  inoculation  in  the  peritoneal  cavity,  in 
producing  the  disease  in  rabbits  and  guinea-pigs,  in  the  form  of 
tumours  of  the  peritoneum.  Sauvageau  and  Radais  consider  that 
actinomyces  should  not  be  included  with  bacteria,  and  have  suggested 
the  name  Oospora  bovis. 

PREPARATION  AXD  EXAMINATION  OF  TISSUES. 

In  order  to  examine  the  microscopical  appearances,  the  tissues  should 
be  hardened  in  absolute  alcohol  and  embedded  in  celloidin.  The  sections,, 
when  stained,  are  to  be  dehydrated  as  a  rule  in  strong  spirit  instead  of 
absolute  alcohol,  as  the  latter  dissolves  the  celloidin.  If  the  sections  are 
very  friable,  they  can  be  cleared  with  clove-oil  on  the  slide.  By  these 
means  the  little  fungus  tufts,  which  have  a  great  tendency  to  fall  out  of 
the  sections,  may  be  preserved  -in  situ  after  passing  through  the  various 
staining  processes.  To  cut  the  sections,  we  can  use  either  Jung's  micro- 
tome, cutting  in  alcohol,  or  the  freezing  microtome.  In  the  latter  case, 
after  the  celloidin  has  hardened,  it  is  necessary  to  shave  off  all  that 
surrounds  the  piece  of  tissue.  It  is  placed  in  water  until  it  sinks,  and 
then  transferred  to  gum,  and  frozen  and  cut  in  the  ordinary  way. 

Staining  Methods. 

There  are  several  methods  by  which  the  organism  can  be  stained  in 
the  tissues,  but  it  is  best  to  employ  for  this  purpose  Gram's  method  and 
modifications  of  Plant's  method. 

Gram's  Method. — By  Gram's  method  the  clubs  in  the  bovine  disease 
are  distinctly  stained,  especially  if  the  sections  contain  the  fungus  at  a 
suitable  stage.  Use  freshly  prepared  staining  solution.  A  few  drops  of 
aniline-oil  are  placed  in  a  test-tube,  which  is  filled  up  with  distilled 
water,  the  mouth  of  the  tube  closed  with  the  thumb,  and  the  mixture 
shaken  up  thoroughly.  An  emulsion  forms,  which  is  then  filtered,  until  a 
perfectly  clear  solution  of  aniline-water  is  obtained.  To  this  is  added, 
drop  by  drop,  an  alcoholic  solution  of  gentian-violet  until  precipitation 
commences.  About  fifteen  to  twenty  drops  in  a  small  capsule  of  aniline- 
water  will  be  sufficient.  Sections  are  floated  in  this  dye  for  about  ten 
minutes,  then  transferred  to  the  iodine-potassic-iodide  solution  until  they 
turn  brown  like  a  tea-leaf.  They  are  then  decolorised  in  alcohol  ;  then 
stained  in  a  weak  alcoholic  solution  of  eosin,  dehydrated  in  strong  com- 
mercial alcohol,  cleared  in  clove-oil,  and  mounted  in  balsam.  It  will  be 
found  that  the  clubs  are  stained  blue,  and  that  there  is  a  central  area, 
which  is,  as  a  rule,  tinged  by  the  eosin.  There  are  various  modifications 
of  the  method,  and  some  of  them  are  extremely  successful  in  affording 
not  only  a  picture  of  the  fungus,  but  also  the  structure  of  the  surrounding 
tissue.  Very  instructive  results  may  be  obtained  by  combining  the 
method  of  Gram  with  Ehrlich's  histological  stain.  In  this  case,  after 
the  section  has  been  decolorised  in  alcohol,  it  is  ready  to  be  transferred 
to  logwood  and  treated  as  described  below. 

Weigert's  Method. — This  also  gives  very  beautiful  results.    The  sections 


440  INFECTIVE   DISEASES. 

are  placed  for  an  hour  in  Wedl's  solution  of  orseille,  which  is  prepared 
as  follows  : — Add  liquid  extract  of  orseille  to  a  mixture  of  absolute 
alcohol  20  parts,  strong  acetic  acid  5  parts,  distilled  water  40  parts, 
until  a  dark-red  liquid  results.  This  must  be  filtered  before  use.  The 
sections  are  left  in  this  solution  for  an  hour,  then  just  rinsed  in  alcohol, 
and  transferred  to  a  solution  of  gentian-violet.  Such  sections  show  the 
nuclei  of  a  violet-blue  colour,  and  the  peripheral  part  of  the  central  core 
in  the  larger  masses  of  the  fungus  also  takes  a  blue  colour,  while  the 
club-shaped  structures  are  stained  a  striking  wine-red  colour. 

Plant's  Method  and  Modifications. — This  is  one  of  the  most  valuable 
methods  for  staining  the  clubs.  The  original  method  was  to  float  sections 
for  ten  minutes  in  magenta  solution  warmed  to  45°  C.  This  solution 
consisted  of  magenta  two  parts,  aniline-oil  3  parts,  alcohol  of  specific 
gravity  0'830,  20  parts,  distilled  water  20  parts  (Gibbes).  The  sections 
were  then  rinsed  in  water,  stained  in  concentrated  alcoholic  solution  of 
picric  acid  for  from  five  to  ten  minutes,  immersed  in  water  five  minutes, 
50  per  cent,  alcohol  fifteen  minutes,  passed  through  absolute  alcohol  and 
clove-oil,  and  preserved  in  Canada  balsam.  The  clubs  are  stained  a 
brilliant  red  and  the  tissue  yellow.  Instead  of  employing  the  magenta 
solution,  we  now  use  Ziehl-Neelsen's  solution. 

By  removing  the  picric  acid  in  Plant's  method  by  prolonged  immersion 
in  alcohol,  and  then  staining  with  gentian-violet  or  methylene-blue,  a 
very  successful  contrast  can  be  obtained.  The  most  instructive  histo- 
logical  picture  can  be  obtained  by  first  staining  with  Neelsen's  solution, 
removing  the  stain  from  the  tissue  in  the  way  that  has  been  already 
described,  and  then  transferring  the  sections  to  distilled  water,  and 
subsequently  staining  with  Ehrlich's  histological  stain. 

Ehrlictis  New  Histological  Stain. — This  is  a  combination  of  Ehrlich's 
logwood  with  orange-rubin.  It  is  of  especial  value  for  sections  of 
actinomycosis,  and  particularly  in  combination  with  carbolised  fuchsine. 
It  is  employed  in  the  following  way  :— The  sections  must  be  placed  in 
alcohol  or  distilled  water,  and  then  in  Ehrlich's  logwood  for  about  half 
a  minute.  From  this  solution  they  are  transferred  to  distilled  water, 
washed  to  remove  the  excess  of  stain,  and  then  placed  in  a  large  dish  of 
tap-water,  where  they  are  left  for  half  an  hour  or  more,  until  the 
sections  turn  blue  ;  if  preferred,  they  may  be  left  overnight.  They  are 
then  stained  for  one  or  two  minutes  in  a  solution  of  rubin,  S,  and 
orange,  and  washed  again  in  distilled  water  to  remove  the  excess.  They 
must  then  be  dehydrated  in  alcohol,  cleared  in  clove-oil,  and  mounted  in 
balsam. 

Preparation  of  Large  Sections. 

The  method  of  cutting  large  sections  of  organs  may  be  employed 
in  studying  actinomycosis.  The  value  of  these  sections  depends  not 
only  upon  their  affording  an  instructive  picture  of  the  naked-eye 
appearances,  but  they  can  also  be  studied  with  a  pocket  lens,  or  under 
the  microscope  with  a  |  or  £-in.  objective.  By  staining  the  sections,  the 
relation  of  the  morbid  to  the  healthy  structures  is  brought  out  in  greater 
contrast,  and  thus  the  topography  of  the  disease  can  be  studied  more 


DESCRIPTION    OF    PLATES    XX.    AND    XXI. 
Actinomycosis  Bovis. 

PLATE    XX. 

FIG.  1.— From  a  section  of  an  actinomycotic  tongue  stained  by  the  triple 
method  (Ziehl-Neelsen,  logwood  and  orange-rubin).  In  this  section  the 
separate  centres  of  growth  are  clearly  shown.  Each  neoplasm  consists  of 
a  fungus  system,  in  which  the  masses  of  the  fungus,  situated  more  or  less 
centrally,  are  surrounded  with  round  cells,  epithelioid  cells,  sometimes 
giant  cells,  and  lastly  fibrous  tissue  forming  a  more  or  less  distinct 
capsule.  In  parts  the  fungi  have  fallen  out  of  the  section.  x  50. 

FIG.  2. — From  a  section  of  a  "tubercular"  nodule  from  the  lungs  of  a 
Norfolk  heifer  with  pulmonary  actinomycosis.  The  nodule  is  a  multiple 
growth  surrounding  a  bronchus,  and  is  enclosed  by  a  capsule,  in  the 
vicinity  of  which  the  pulmonary  alveoli  are  compressed.  It  is  composed 
of  a  number  of  separate  neoplasms,  and  each  of  the  latter  is  composed  of 
secondary  centres  of  growth  resembling  the  giant  cell  systems  of  bacillary 
tuberculosis.  The  new  growth  is  composed  of  ray-fungi,  large  multi- 
nucleated  cells,  sometimes  distinct  giant  cells,  round  cells,  epithelioid  cells, 
and,  surrounding  them,  fibrous  tissue.  On  examination  of  the  same 
specimen  with  a  higher  power  the  typical  rosettes  of  clubs  are  sometimes 
surrounded  by  multinucleated  cells,  and  sometimes  small  rosettes  are 
found  like  tubercle  bacilli,  in  the  interior  of  giant  cells.  From  a  pre- 
paration stained  by  Ziehl-Neelsen,  logwood,  and  orange-rubin.  x  50. 

PLATE  XXI. 

FiG.  1. — (a)  A  leucocyte  containing  the  fungus  in  its  earliest  recognisable 
form.  (&)  A  large  multinucleated  cell  containing  the  fungus  in  an  early 
stage  with  the  club-form  already  visible,  (c)  A  leucocyte  containing  a 
small  stellate  fungus,  (d)  A  large  cell  containing  clubs  arranged  in  a 
small  rosette.  (<?)  A  multinucleated  cell  with  clubs  arranged  in  a  palmate 
form.  All  the  above  are  drawn  from  sections  of  actinomycotic  tongues 
stained  by  the  triple  method,  x  500. 

FIG.  2. — A  giant  cell  with  large  vesicular,  nuclei  at  the  periphery,  and  in  the 
centre  a  fully  formed  rosette  of  actinomyces  with  a  smaller  growth  within 
a  "  daughter "  cell.  From  a  section  of  the  tongue  of  an  ox  stained  by 
the  triple  method,  x  500. 

FIG.  3. — A  very  large  circular  giant  cell,  with  its  ring  of  nuclei  at  the 
periphery,  enclosing  several  isolated  tufts  of  actinomyces.  From  a  section 
of-  a  nodule  in  the  lung.  Stained  by  the  triple  method,  x  500. 

FIG.  4. — Three  rosettes  of  actinomyces  surrounded  by  a  row  of  large,  some- 
what angular  multinucleated  cells.  From  a  section  of  the  tongue  of  an 
ox  stained  by  the  triple  method,  x  430. 


^^-;*^^>^^v>'     •-  "T^H'^wsw 


-^r 


ruwUc/eca. 


.CTINOMYCOSIS  B' 


DESCRIPTION    OF    I'J  I)    XXI. 

Actinomycosis  Bovia. 

PL  A 

FlG.  1. — From  a  section  of  an  acth  tined  by  the  triple 

method  (Ziehl-Neelsen,  logwo«>  abin).  In  this  section  the 

separate  centres  of  growth  are  i  i  neoplasm  consists  of 

a  fungus  system,  in  which  the  mass-  s,  situated  more  or  less 

centrally,  are  surrounded  lioid  cells,  sometimes 

giant  cells,  and  lastly  f  more  or  less  distinct 

capsule.  In  parts  the  i  otlon.  x  50. 

FIG.  2. — From  a  section  of  nodule  from  the  lungs  of  a 
Norfolk  heifer  with  puhn  The  nodule  is  a  multiple 
growth  surrounding  »sed  by  a  capsule,  in  the 
vicinity  of  which  tin  ompressed.  It  is  composed 
of  a  number  of  separ  .if  the  latter  is  composed  of 
secondary  centres  of  nt  cell  systems  of  bacillary 
tuberculosis.  The  new  L  of  ray-fungi,  large  multi- 
nucleated  cells.  i -11s.  round  cells,  epithelioid  cells, 
and,  surrounding  r  liination  of  the  same 
specimen  with  a  higher  power  ;  if  clubs  are  sometimes 
surrounded  by  innh  M it-times  small  rosettes  are 
found  like  tub-  :->r  of  giant  cells.  From  a  pre- 
paration stained  M  :u<l  «»range-rubin.  x  50. 

FIG.    !  -us  in  its  earliest  recognisable 

utaining  the  fungus  in  an  early 
(c)  A.  leucocyte  containing  a 

smai  .lining  clubs  arranged  in  a 

smnli  •  lubs  arranged  in  a  palmate 

form.     A  of  actinomycotic  tongues 

i  at  the  periphery,  and  in  the 

smaller  growth  within 

of  an  ox  stained  by 

thr  : 

FIG.  ::ig  of  nuclei  at  the 

peri]  •  :iyces.     From  a  section 

of;  triple  method,      x  500. 

by  a  row  of  large,  sorne- 
From  a  section  of  the  tongue  of  an 
•:uned  b\  T:  *    l.iO. 


Plate  XXI. 


1 


^ 


* 


Pig  4 


ACTINOMYCOSIS  BO^IS 


'Vincent  Bno1ej,Dty  4Son,UtK . 


442  INFECTIVE   DISEASES. 

of  best  filter- paper.  In  this  way  several  large  sections  can  be  cleared  at 
the  same  time.  But  when  only  one  or  two  sections  are  dealt  with,  they 
are  cleared  in  clove-oil  in  a  dish,  and  the  mounting  glass  at  this  stage 
passed  underneath  them  as  already  described.  Another  plan,  which  will 
be  found  of  advantage,  is  as  follows  : — A  piece  of  clean,  thick,  filter-paper 
rather  larger  than  the  section,  is  slipped  underneath  it,  and  then  raised 
with  the  section  upon  it.  After  allowing  the  excess  of  clove-oil  to  drain 
back  into  the  dish,  it  is  carefully  laid  on  the  glass  with  the  section  down- 
wards, and  gently  pressed  down.  By  taking  up  a  corner,  the  filter-paper 
is  peeled  off,  and  the  section  left  behind  on  the  glass.  Any  creases  or 
folds  are  adjusted  with  needles.  After  removal  of  the  clove-oil,  balsam 
is  run  over  the  section,  and  a  cover-glass  gently  and  dexterously  lowered, 
so  as  to  avoid  the  presence  of  air-bubbles.  The  preparations  are  set  aside 
to  harden  in  a  warm  place  and  on  a  level  surface,  and  are  then  ready  for 
fixing  in  suitable  frames. 

Naked-eye  Appearances  of  Large  Sections. 

In  the  sections  of  an  actinomycotic  tongue  it  is  at  once  apparent  that 
the  new  growth  is  more  or  less  limited  to  the  periphery  of  the  section . 
In  .parts  there  are  dense  clusters  of  little  nodular  neoplasms,  the  fungus 
syxtems,  each  having  a  rounded  form,  and  averaging  in  size  that  of  a 
small  pea.  In  other  parts  small  nodules,  varying  in  size  from  a  millet- 
seed  to  a  hemp-seed,  have  a  linear  arrangement  between  bundles  of 
muscular  fibres.  The  appearance  is  suggestive  of  an  invasion  of  the 
tongue  along  the  lymphatics. 

In  many  of  the  nodules  the  largest  tufts  of  the  fungus  can  be  seenr 
with  the  naked  eye,  to  occupy  a  more  or  less  central  position.  In  parts 
the  muscular  fibres  are  replaced  by  fibrous  tissue. 

If  now  these  sections  be  placed  under  the  microscope,  the  minute 
structure  may  be  examined  ;  but  as  it  is  obvious  that  still  better  results 
may  be  obtained  by  small  sections,  any  part  which  it  is  necessary  to 
examine  with  high  powers  can  be  selected  from  a  corresponding  part  of 
the  growth,  and  prepared  in  the  ordinary  way. 

In  the  case  of  a  "  wen,"  the  whole  growth  can  be  excised  with  the 
surrounding  tissues,  sliced  and  treated  in  the  way  already  described,  and 
sections  stained  by  different  methods. 

The  nature  of  the  growth  is  at  once  recognisable  as  aetinomycosis, 
from  the  characteristic  honeycombed  appearance  produced  by  the  trabeculae 
of  fibrous  tissue  which  form  a  spongy  structure,  from  the  loculi  of  which 
the  fungus  tufts  and  caseous  matter  have  for  the  most  part  dropped  out. 
In  other  parts  this  structure  is  intact,  and  the  tufts  of  the  fungus  can  be 
detected  with  the  naked  eye,  and  readily  recognised  with  a  pocket  lens. 

A  fungus  system  may  be  studied  more  minutely  in  ordinary  sections 
of  the  tongue.  Each  nodule  is  composed  of  the  actinomyces  surrounded 
by  round  cells  and  epitheloid  cells,  and  fibrous  tissue  which  often  forms 
a  distinct  capsule.  In  some  specimens  the  fungus  is  surrounded  by  a 
single  row  of  large  multinucleated  cells,  and  in  other  specimens  the 
fungus  is  found  in  the  interior  of  large  oval  giant-cells. 


ACTINOMYCOSIS.  443 

TRANSMISSION  OF  ACTINOMYCOSIS  FROM  MAN  TO  THE  LOWER  ANIMALS. 

Inoculation  of  cultures  has  already  been  referred  to.  The 
author  successfully  inoculated  a  calf  with  material  direct  from 
a  living  patient. 

A  calf  which  had  been  inoculated  in  the  peritoneal  cavity,  and 
killed  seventy  days  afterwards,  presented  the  following  lesions.  The 
peritoneum  of  the  rumen,  in  the  vicinity  of  the  seat  of  inoculation, 
was  studded  with  hundreds  of  growths,  varying  in  size  from  a  millet- 
seed  to  a  pea.  The  large  growths  were  composed  of  several  small 
ones  collected  together.  On  stripping  off  the  peritoneum,  and  holding 
it  between  the  light  and  the  eye,  the  fungus  could  be  seen  with 
the  naked  eye  in  each  individual  growth.  By  incising  a  growth, 
and  examining  a  scraping  under  the  microscope,  the  characteristic 
clubs,  and  the  filaments  also,  were  found  to  be  present.  By  staining 
cover-glass  preparations  with  the  method  of  Gram  and  orange-rubin, 
the  appearances  were  very  striking.  The  clubs  were  conspicuous  on 
account  of  their  size,  and  brilliantly  stained.  In  many,  the  proto- 
plasm of  the  thread  was  demonstrated  in  the  interior  of  the  club. 
In  sections  of  the  peritoneal  nodules  stained  by  Gram's  method 
the  mycelium  was  found  to  be  present,  and  the  clubs  in  part 
took  the  stain.  With  Plant's  method  the  clubs  were  most  clearly 
demonstrated. 

Israel  and  Johne  failed  to  infect  a  calf  by  intravenous  injec- 
tion, and  Ponfick  failed  to  infect  dogs,  but  Israel  succeeded  with 
a  rabbit.  Israel  obtained  a  small  piece  of  actinomycotic  granula- 
tion tissue  from  a  peri-pleural  abscess  in  a  patient  with  primary 
disease  of  the  lung,  and  introduced  it  into  the  peritoneal  cavity. 
The  rabbit  showred  no  sign  of  illness,  and  was  killed  about  ten 
weeks  afterwards.  On  examination  numbers  of  tumours  were 
found  in  the  abdominal  cavity,  varying  in  size  from  a  hemp-seed 
to  a  cherry.  Larger  ones  had  a  somewhat  nodular  surface  with 
yellowish  points ;  others  were  found  on  the  abdominal  wall  011 
the  right  side.  There  was  a  small  growth  over  the  psoas  muscle, 
Miid  one  large  one  attached  to  an  adhesion  of  the  colon.  The 
growths  were  on  the  peritoneum,  or  attached  by  longer  or  shorter 
adhesions.  Some  of  the  larger  tumours  showed  on  section  a  hollow 
space  in  the  centre,  wrhich  was  filled  with  a  pulp,  the  result  of 
fatty  degeneration  of  the  transplanted  tissue,  which  was  sharply 
differentiated  in  colour  and  consistency  from  the  new  growths.  The 
latter  consisted  of  granulation  tissue  with  abundant  formation  of 
fat  granules,  blood  pigment,  acicular  fat  crystals,  and  actinomycotic 


444  INFECTIVE   DISEASES. 

grains.  From  some  of  the  tumours,  radiating  processes  of  a  yellowish 
colour  penetrated  into  the  retro- peritoneal  tissue. 

Rotter  inoculated  calvas,  pigs,  guinea-pigs,  and  rabbits.  In  one 
case  he  had  a  positive  result.  A  piece  of  actinomycotic  growth  was 
introduced  into  the  peritoneal  cavity  of  a  rabbit.  The  animal  was 
killed  six  months  afterwards,  and  the  piece  of  tissue  which  had  been 
introduced  was  found  to  be  encapsuled,  arid  around  it  were  twenty 
tumours,  from  the  size  of  a  pin's  head  to  that  of  a  hazel-nut, 
containing  the  ray- fungus. 

The  author  also  succeeded  in  transmitting  the  disease  to  a  rabbit. 
A  small  quantity  of  human  pus,  containing  the  yellow  grains,  was 
diffused  in  broth,  and  injected  with  a  hypodermic  syringe  into  the 
abdominal  cavity  of  a  rabbit.  This  rabbit  died  seventy- nine  days 
afterwards.  On  examination  several  nodules  were  found,  about  the 
size  of  a  millet-seed,  on  the  peritoneum  of  the  stomach,  in  the  gastro- 
splenic  omentum,  and  on  the  peritoneum  of  the  diaphragm.  There 
was  a  rounded  nodule,  about  the  size  of  a  pea,  attached  to  the 
stomach.  There  were  adhesions  between  the  intestines,  and  a 
tumour  about  the  size  of  a  marble  attached  to  an  adhesion  of 
the  caecum.  One  of  the  small  nodules  was  excised  and  divided,  and 
the  scraping  from  the  interior  contained  typical  rosettes  of  clubs. 

The  successful  transmission  of  actinomycosis  from  man  to  bovines 
suggests  intercommunicability,  though  the  negative  evidence  as  to 
infection  of  man  from  bovines  supports  the  view  that  the  disease 
is  derived  from  some  source  which  is  common  to  both  species. 

TRANSMISSION  OF  ACTINOMYCOSIS  FROM  CATTLE  TO  CATTLE. 

Johne  was  the  first  to  prove  that  actinomycosis  could  be  trans- 
mitted from  cattle  to  cattle,  and  his  results  were  confirmed  and 
extended  by  Ponfick. 

A  calf  was  inoculated  subcutaneously  in  the  neck  and  cheek, 
in  the  gum,  and  the  abdominal  cavity.  The  animal  died  in  forty 
days  after  inoculation  with  development  of  actinomycosis. 

A  calf  was  inoculated  in  the  cheek  and  abdominal  cavity.  Death 
occurred  114  days  after  inoculation.  In  the  peritoneal  cavity 
numerous  tumours  had  formed,  and  the  yellowish  grains  were  visible 
to  the  naked  eye  in  sections  of  the  new  growth. 

A  cow  in  calf  was  inoculated  in  the  left  posterior  quarter  of  the 
udder ;  phlegmonous  mastitis  followed,  and  subsided  leaving  a  small 
induration,  which  then  increased  until  the  inoculated  part  of  the 
udder  was,  in  three  months,  nearly  double  the  normal  size,  from  a 


ACTINOMYCOSIS.  ,  445 

deposit  resembling  a  fibroma.  The  cow  was  slaughtered  133  days- 
after  inoculation.  Typical  actinomycosis  had  been  produced. 

A  colt  three  and  a  half  years  old,  was  inoculated  in  the  jaw 
and  in  the  forehead  after  trephining,  and  also  in  the  trachea.  The 
animal  died  without  any  result. 

Ponfick  also  conducted  a  series  of  experiments  which  amply 
confirmed  the  results  which  had  been  obtained  by  Johne. 

Feeding  Experiments. — Repeated  experiments,  made  with  masses- 
of  the  growth  chopped-  up,  or  with  isolated  grains  of  the  fungus,  gave 
negative  results. 

Inoculation  Experiments. — The  growth  was  inoculated  in  various 
regions  of  the  body.  Small  particles  of  the  growth  from  quite  fresh 
tumours  were  introduced  into  the  anterior  chamber  of  the  eye  in 
rabbits,  with  negative  results.  Rabbits  were  inoculated  in  the 
peritoneal  cavity  from  an  animal  recently  slaughtered,  but  they 
died  of  peritonitis.  In  dogs  also  the  results  were  negative. 

Seven  calves  were  operated  on.  In  five  the  abdomen  was  opened 
with  antiseptic  precautions,  and  in  two  cases  the  growth  was 
introduced  by  injection.  In  the  latter  cases  the  pieces  of  tumour 
were  suspended  in  salt  solution,  but  the  animals  died  from  peritonitis 
a  few  days  after  the  injection.  The  same  result  occurred  in  two  out 
of  the  five  cases  in  which  the  abdomen  was  opened.  The  three 
remaining  cases  gave  the  following  results  : — 

1. — Pieces  of  tissue,  about  1*5  cm.  long,  were  taken  from  the 
lower  jaw  of  a  recently  killed  ox.  Twelve  of  these  pieces  were 
introduced  into  the  peritoneal  cavity;  death  occurred  after  266 
days,  from  exhaustion  and  recent  lobular  pneumonia.  At  the  post- 
mortem examination  several  patches  of  peritonitis  were  found,  with 
encystment  of  the  remains  of  fragments  of  the  inoculated  tissue, 
but  there  was  an  independent  development  of  several  nodules  in  the 
neighbourhood  of  the  stomach  and  urinary  bladder.  Examination 
of  these  new  formations  showed,  even  to  the  naked  eye,  that  they 
contained  yellowish  grains,  which,  on  further  examination,  proved 
to  be  the  fungi. 

2. — Ten  pieces  of  tumour  from  the  jaw  of  a  cow  were  introduced 
as  before ;  the  calf  died  suddenly  sixty  days  afterwards,  during  the 
injection  of  fresh  pieces  of  actinomycosis  into  the  jugular  vein.  At 
the  post  mortem  it  was  found  that  various  adhesions  had  occurred, 
as  the  result  of  peritonitis ;  and  in  the  false  membranes  there  were 
sixty-three  nodules,  varying  in  size.  Microscopical  examination 
showed  that  all  these  nodules  consisted  of  typical  actinomycotic  new 
formation. 


446  INFECTIVE   DISEASES. 

3 in  this  case  twelve  pieces  of  growth   were  introduced  into 

the  abdomen  of  a  calf  eight  weeks  old.  Seven  days  afterwards  fresh 
pieces  were  introduced  under  the  skin,  in  the  region  of  the  left  lower 
jaw.  A  swelling  occurred,  which  was  opened.  Pus  escaped, 
together  with  the  material  which  had  been  used  for  inoculation,  in 
a  state  of  decomposition.  Pieces  of  tumour  were  inoculated  sub- 
cutaneously  in  the  neighbourhood  of  the  right  side  of  the  neck, 
ninety-nine  days  after  the  first  experiment.  The  animal  was  bled  to 
death  seven  months  (210  days)  after  the  first  experiment.  At  the 
post  mortem,  peritonitis  and  adhesions  were  found,  with  twenty-one 
large  and  several  small  nodules  in  the  mesentery,  in  the  false  mem- 
branes between  the  viscera,  and  on  and  in  the  serous  linings  of  most 
of  the  abdominal  organs.  There  were  also  several  large  and  many 
.small  tumours  in  the  subcutaneous  and  inter  muscular  tissue,  in  the 
region  of  the  lower  jaw  and  neck  on  the  right  side,  and  numerous 
large  and  small  nodules  in  both  lungs,  some  undergoing  softening 
in  the  centre. 

Isolated  fungi  were  inoculated  in  dogs,  with  negative  results, 
nothing  remaining  after  45  to  80  days,  except  a  thick  emulsion. 

Pieces  of  tumour  were  introduced  in  the  submucous  and  sub- 
cutaneous tissue  of  dogs,  but  no  change  occurred  after  600  days. 

In  a  calf,  inoculation  under  the  gum  of  the  upper  jaw  showed 
what  was  possiblv  only  the  remains  of  the  growth  which  had  been 
introduced.  At  any  rate,  the  experiment  was  doubtful  ;  but  in 
a  second  calf  there  were  numbers  of  nodules  developed  around 
the  points  of  inoculation  in  the  subcutaneous  and  muscular  tissue 
in  the  neighbourhood  of  the  lower  jaw,  and  in  the  region  of  the 
neck.  The  results  were  observed  in  these  regions  in  210  and  110 
days  respectively. 

Inoculation  of  rabbits  and  dogs,  with  isolated  fungi,  produced 
no  results  after  156,  165,  and  170  days.  Experiments  on  dogs, 
with  grains  from  a  human  source,  were  also  unsuccessful.  They 
were  examined  after  470  days,  and  there  was  no  sign  of  any 
result. 

After  intravenous  injection,  positive  results  were  discovered  m 
the  only  calf  which  survived  this  operation,  on  bleeding  the  animal 
to  death  110  days  afterwards.  Numerous  nodules  (27)  were  dis- 
covered in  the  parenchyma  of  both  lungs,  without  suppuration. 

Two  dogs  were  injected  in  the  jugular  vein  with  isolated  fungi 
mixed  with  60  grammes  of  salt  solution.  Examined  after  45  arid 
80  days  respectively,  the  lungs  and  all  the  other  organs  were  found 
to  be  free  from  growths. 


ACTINOMYCOSIS.  447 

Ponfick  thus  summarised  these  experiments  with  the  bovine 
fungus  : — 

1.  Rabbits  and  dogs  possess  a  marked  immunity  from  actiiio- 
mycosis,  whether  pieces  of  tumour  or  isolated  grains  are  administered 
by  feeding,  or  by  inoculation  in  the  serous  cavities,  in  the  subcutaneous 
or  submucous  tissue,  or  by  intravenous  injection. 

2.  The  most  common  subject  of  actinomycosis,  the  cow,  possesses 
a  not  less  marked  susceptibility  to  the  artificial  production  of  the 
disease.       By   feeding,   an    infection   was    not   obtained,    probably 
because   the    mucous    membrane    had    not   been   injured ;    but   by 
inoculation,    on    the    contrary,    an    independent    growth    of    fresh 
neoplasms   was  produced  in   the  subcutaneous   and   intermuscular. 
tissues,    occasionally   in   the   submucous    tissue,    and   in    a   decided 
manner  in  the  abdominal  cavity.     Clear  evidence  of  this   growth 
is    obtained   in    some    cases    within   a    month,    or   after   three   or 
four  months. 

3.  By  intravenous  injection,  also,  it  is  possible  in  a  few  months 
to  cause  typical  new  growths  in  the  lungs. 

MADURA  DISEASE. 

Mycetoma,  or  Madura  foot,  is  a  chronic  local  disease,  attacking 
<-hiefly  the  hands  and  feet,  and  having  considerable  resemblance  to 
actinomycosis.  It  is  a  disease  of  tropical  climates,  and  is  commonly 
known  as  the  "  fungus-foot  disease  "  of  India.  A  small  tumour  forms 
on  the  hand  or  foot,  which  after  a  year  or  two  suppurates  and 
bursts,  leaving  one  or  more  sinuses,  from  which  peculiar  black 
particles,  or  white  or  pinkish  roe-like  bodies,  are  discharged. 

The  disease  in  the  foot  may  commence  in  the  big  toe  and  spread 
upwards,  involving  the  leg  as  far  as  the  knee,  and  even  the  thigh. 
In  a  typical  case  the  foot  is  enlarged  and  painful,  and  later  there 
are  several  sinuses  from  which  a  purulent  and  blood-stained  discharge 
can  be  expressed,  containing  the  characteristic  particles. 

According  to  Bocarro  all  early  growths  are  superficial.  Dissec- 
tion of  the  growths  during  an  operation,  or  sections  made  through 
the  diseased  tissues  after  excision  or  amputation,  show  that  the 
disease  begins  generally  in  the  loose  cellular  tissue,  generally  the 
subcutaneous  tissue,  and  thence  extends  along  the  sheath  of  muscles 
and  tendons  to  other  soft  tissues,  and  finally  the  bones. 

There  are  several  facts  in  connection  with  the  causation  of  the 
disease,  which  are  of  great  interest  when  it  is  compared  to  actino- 
mycosis in  cattle.  Bocarro  states  that  the  disease  originates  in 


448 


INFECTIVE   DISEASES. 


wounds,  sores  and  pricks  of  thorns,  and  that  the  points  of  the 
thorns  of  the  Acacia  Arabica  have  been  found  embedded  in  the 
diseased  parts.  The  disease  is  common  among  the  agricultural 
class,  and  in  90  per  cent,  of  the  cases  observed  in  the  Hyderabad 
Civil  Hospital  it  occurred  in  the  hands  and  feet. 

Vandyke  Carter  was  the  first  to  point  out  the  resemblance  to 
actinomycosis,  and  he  believed  that  the  two  varieties  of  the  disease, 
the  black  and  the  white,  were  the  result  of  the  growth  of  a  mycelial 
fungus,  Chionyphe  Carteri. 

Kanthack  pointed  out,  that  if  portions  of  the  growth  were 
placed  in  ether  or  chloroform,  and  afterwards  well  washed  in 


FIG.  183. — PAET  OP  HUMAN  FOOT  WITH  MADURA  DISEASE. 

caustic  potash,  small  rounded  bodies  were  left,  which  showed  rays 
under  the  microscope  closely  resembling  the  appearances  in  actino- 
mycosis, and  that  the  reaction  of  the  fungus  to  staining  reagents 
was  identical  with  actinomyces.  Hewlett  examined  sections  from 
the  disease  in  the  foot,  and  also  found  filaments  and  clubs. 
Boyce  and  Surveyor  examined  a  number  of  cases,  and  care- 
fully studied  the  fungus  in  the  black  and  white  varieties  of 
the  disease.  In  the  black  variety  the  particles  were  found  to 
vary  greatly  in  size,  from  that  of  a  grain  of  gunpowder  to  that 
of  a  marble.  If  the  particles  were  boiled  for  from  a  few  minutes 
to  one  hour  in  concentrated  caustic  potash,  and  then  transferred 
to  distilled  water,  the  brown  colouring  matter  was  removed  and  a 


MADURA   DISEASE.  449 

mycelial  fungus  could  be  seen.  If  tissue,  containing  particles,  was 
washed  for  about  a  minute  in  eau  de  Javel  and  then  stained,  the 
colouring  matter  was  removed,  and  the  relation  of  the  fungus  to 
the  tissue  could  be  observed.  This  fungus  consisted  of  large  radiating 
and  branched  hyphse,  like  those  of  a  species  of  aspergillus,  or  mucor. 
Sections  of  the  white,  fish-roe  bodies  showed,  usually  in  the  centre, 
numerous,  small,  reniform,  deeply-stained  masses,  surrounded  by 
a  radiated  zone,  with  the  presence  of  dwarfed  club-like  elements 
resembling  actinomyces. 

The  author  suggested  that  possibly  the  presence  of  the  coarser 
septate  mycelium  of  the  black  variety  might  be  attributed  to  a 
mixed  infection. 

Vincent  in  Algiers  succeeded  in  cultivating  the  micro-organism, 
and  showed  that  it  was  a  new  species  of  streptothrix. 

Streptothrix  madurse. — Yincent  found  that  the  streptothrix 
at  first  grows  scantily  in  the  ordinary  culture  media,  and  in  such 
liquids  as  Cohn's  solution.  In  broth,  at  the  end  of  about  a  fortnight, 
there  is  a  limited  growth  composed  of  small,  round,  greyish  masses, 
and  in  sub-cultures  the  growth  becomes  more  abundant. 

The  little  colonies  float  in  the  clear  liquid  when  the  tube  is 
shaken,  and  subside  to  the  bottom  when  the  liquid  is  at  rest,  while 
some  adhere  to  the  sides  of  the  tube.  They  may  be  very  small,  or 
attain  the  size  of  a  pea  ;  after  two  months  they  acquire  a  reddish 
tinge.  Later,  on  the  surface  of  the  liquid,  there  is  a  white  efflores- 
cence composed  of  spores. 

The  streptothrix  grows  well  in  slightly  acid  infusions  of  hay  or 
straw,  the  proportion  of  hay  to  water  being  15  grammes  to  the 
litre.  Vegetable  infusions,  made  with  carrots,  turnips,  and  potatoes 
(20  grammes  to  1,000  of  water),  are  suitable  media.  The 
streptothrix  grows  at  the  temperature  of  the  room,  but  best  at  37°  C. 
and  with  free  access  to  air.  Inoculated  in  the  depth  of  gelatine  there 
is  a  scanty  growth  in  the  track  of  the  needle  and  on  the  surface ; 
but  it  grows  best  in  a  nutrient  medium,  composed  of  infusion  of 
hay  or  potato  100  cc.,  gelatine  9  grammes,  glycerine  4  grammes, 
and  grape-sugar  4  grammes.  Gelatine  is  not  liquefied. 

Ordinary  nutrient  agar  is  not  a  very  favourable  medium,  but 
on  glycerine-agar  with  grape-sugar  there  is  an  abundant  growth  of 
circular,  projecting,  shining  colonies,  slightly  yellowish -white,  which 
later  become  pink  or  bright  red.  When  the  colonies  are  numerous 
they  remain  small,  but  isolated  colonies  increase  rapidly  ;  they  are 
depressed  in  the  centre  or  umbilicated,  and  the  central  part  remains 
white  while  the  periphery  becomes  red.  Later,  the  culture  loses  its 

29 


450  INFECTIVE   DISEASES. 

colour  and  becomes  a  dull  white.  The  growth  is  very  adherent  to 
the  surface  of  the  jelly,  and  so  tough  that  it  is  almost  horny.  The 
Streptothrix  can  be  cultivated  in  milk,  which  it  slowly  pepto- 
nises.  It  cannot  be  grown  on  blood  serum  or  egg.  On  potato,  on 
the  fifth  day  at  37°  C.,  there  are  little  prominent  colonies,  which 
.slowly  increase.  After  a  month  the  growth  acquires  a  pale-rose 
colour,  which  gradually  increases  and  changes  to  orange  or  dark 
red.  The  colour  is  most  intense  on  acid  potato,  and  after  a  time 
an  efflorescence,  or  whitish  dust,  appears  on  some  of  the  colonies, 
consisting  of  spores.  The  growth  is  hard  and  friable. 

Rabbits,  mice,  guinea-pigs,  and  a  cat,  were  inoculated  sub- 
cutaneously  with  particles  from  the  disease,  or  with  cultures ;  but 
only  a  local  nodule  was  produced  in  each  case,  which,  after  a  slight 
increase,  subsided.  Nocard  confirmed  these  results.  Intra-peritoneal 
;and  intravenous  and  subcutaneous  inoculations  in  guinea-pigs,  rabbits, 
pigeons,  fowls,  dogs,  and  sheep  were  negative ;  and  no  trace  could 
be  found  of  the  cultures  in  any  animal  subsequently  killed  and 
•examined.  According  to  Bocarro,  though  fresh  particles  from  the 
disease  inoculated  in  rabbits  and  dogs  gave  negative  results,  inocu- 
lation of  cultures  in  guinea-pigs,  rabbits,  monkeys,  and  rats,  pro- 
duced a  local  tumour  of  slow  growth,  which,  on  section,  had  the 
character  of  the  inoculated  material. 

From  these  experiments  we  must  conclude  that  the  disease  has 
not  been  shown  to  be  transmissible  to  the  lower  animals,  by  either 
inoculation  of  the  diseased  tissue,  or  by  cultures  of  the  Streptothrix  ; 
and  the  exact  relation  of  both  the  Streptothrix  and  the  mycelial 
fungus  to  the  disease  must  be  considered  an  open  question. 


CHAPTEE  XXXI. 

GLANDERS. 

GLANDERS  is  a  specific  inocalable  disease  of  equines,  characterised 
by  the  formation  of  nodules  and  suppurating  tumours,  with  which 
characteristic  bacilli  are  associated.  The  disease  has  been  known 
from  very  early  times.  It  is  described  in  books  of  the  sixteenth 
century  and  in  very  early  treatises  on  farriery.  It  attacks  horses, 
asses,  and  mules.  Man  and  many  of  the  lower  animals  can  be 
readily  inoculated,  but  cattle  and  swine  have  an  immunity.  The 
disease  is  especially  prevalent  in  towns,  or  wherever  horses  may  be 
crowded  together  without  those  sanitary  arrangements  which  are 
so  much  attended  to  in  private  stables ;  and  in  large  establishments 
fresh  horses  are  being  constantly  introduced  to  replace  others,  and 
thus  the  opportunities  for  the  importation  of  the  disease  are  multi- 
plied. The  disease  varies  in  its  virulence.  It  may  occur  in  a  form 
which  proves  fatal  in  a  few  days,  or  it  may  exist  for  months  or 
years  without  attracting  notice,  and  yet  be  capable  of  being  trans- 
mitted to  other  animals.  The  term  "farcy"  is  applied  when  the 
disease  manifests  itself  in  the  form  of  tumours  in  the  skin. 

Glanders  in  the  horse  most  commonly  produces  ulcer ation  of  the 
nostrils  and  enlargement  of  the  glands.  It  commences  in  the  form 
of  nodules  of  the  mucous  membrane  resembling  miliary  tubercles, 
and,  like  them,  consisting  of  a  collection  of  round  cells.  They 
suppurate  and  coalesce,  forming  irregular  ulcers  and  raised,  congested 
nodules.  The  lymphatic  glands  become  enlarged  and  suppurate, 
and  the  disease  extends  to  the  respiratory  organs.  In  the  lungs,  in 
the  early  stage,  the  disease  is  readily  mistaken  for  tuberculosis. 
The  nodules  suppurate,  and  cavities  are  formed,  but  they  do  not 
tend  to  caseate.  A  glairy  or  muco- purulent  discharge  from  the 
nostrils  should  lead  to  very  careful  inspection,  with  every  possible 
precaution ;  and  it  will  probably  be  easy  to  detect  the  ulceration  of 
the  nostrils.  In  other  cases  there  may  be  slight  discharge  from  the 

451 


452 


INFECTIVE   DISEASES. 


nostrils  and  swelling  of  the  glands,  and  nothing  more  will  be  visible 
until  a  post  mortem  examination  has  been  made. 

Glanders  in  man  is  found  amongst  those  whose  duties  bring 
them  into  contact  with  diseased  horses,  such  as  stable-men,  cavalry 
soldiers,  and  veterinary  surgeons ;  and  it  is  generally  the  result 
of  accidental  inoculation  of  a  wound.  An  abscess  develops,  followed 
by  metastatic  purulent  infiltration  of  the  lungs,  liver,  spleen,  and 
bones.  There  may  be  cedeinatous  swelling  of  the  face  and  ulcers 
in  the  nostrils.  The  joints  may  become  swollen  and  painful. 

The  nodules  consist  of  fibrous  tissue  and  cells,  with  a  tendency  to 
suppuration.  In  the  lungs  the  disease  spreads  by  the  lymphatics. 


iff; 

*££. 
ife«r^ 


FIG.  184.— BACILLI  OP  GLANDERS  ;  section  of  a  glanders  nodule,  x  700  (FLUGGE).. 


The  infiltrated  patches  are  necrosed  in  the  centre,  which  is  sur- 
rounded by  leucocytes  and  fibrous  tissue. 

The  bacilli  were  discovered  by  Loffler  and  Schutz  in  1882.  They 
are  found  in  the  discharge  from  the  nostrils,  in  the  pus,  and  in  the 
nodules  of  animals  artificially  infected. 

Bacillus  mallei — Rods,  with  rounded  ends,  shorter  and 
thicker  than  the  .tubercle  bacillus,  occurring  singly,  or  in  pairs, 
and  sometimes  in  filaments.  The  protoplasm  in  the  rod  is  broken 
up  in  stained  preparations,  as  in  the  tubercle  bacillus.  They  stain 
with  the  watery  aniline  dyes,  and  intensely  so  with  alkaline  methylene 
blue  or  Neelsen's  solution ;  they  are  non-motile  and  aerobic  ;  spore 
formation  has  been  described.  They  can  be  cultivated  on  the  usual 
media,  especially  on  glycerine-agar  and  on  potato ;  but  they  will  not 
grow  in  infusions  of  hay,  straw,  or  stable  manure.  On  the  surface 
of  glycerine-agar  a  colourless,  transparent  growth  occurs  on  either 


GLANDERS. 


4,53 


side  of  the  track  of  the  needle;  on  glycerine-agar  with  milk  a 
whitish  layer  develops,  which  gradually  changes  in  colour  from 
amber  yellow  to  a  reddish-brown.  On  blood  serum  the  growth  is 
transparent  and  yellowish  ;  on  potato  it  is  much  more  characteristic. 
After  two  or  three  days  at  the  temperature  of  the  blood,  a  film 
develops  in  the  vicinity  of  the  inoculated  area,  which  is  honey-like, 
transparent  and  yellow ;  the  transparency  disappears,  and  in  a 
week  the  cultures  have  become  reddish-brown.  (Plate  II.,  Fig.  6.) 

The  disease  has  been  communicated  to  man  by  accidental 
inoculation  with  a  hypodermic  syringe  which  had  been  used  for 
inoculating  cultures.  Horses,  asses,  cats,  goats,  field  mice,  and 


FIG.  185.— SECTION  OF  A  BRANCH  OF  THE  PULMONARY  ARTERY  SHOWING 
GLANDERS  BACILLI  PENETRATING  THE  WALL  (HAMILTON). 

guinea-pigs,  can  all  be  infected  with  pure  cultures ;  rabbits,  sheep, 
and  dogs  are  slightly  susceptible;  cattle,  swine,  and  white  mice 
have  an  immunity.  In  the  guinea-pig  a  swelling  occurs  at  the  seat 
of  inoculation,  followed  by  the  formation  of  a  prominent  tumour, 
developing  into  an  abscess.  The  skin  becomes  involved,  and  an  ulcer 
with  indurated  margin  results.  The  lymphatic  glands  also  become 
implicated,  and  general(  infection  follows,  extending  frequently  to 
the  testicles  or  ovaries,  and  death  results  after  several  weeks.  In 
rabbits  there  is  generally  only  a  local  abscess  induced,  which  termi- 
nates in  a  quickly-healing  sore.  Mice  die  in  three  or  four  days 
from  general  infection ;  glanders  nodules  are  found  in  the  liver  and 


454  INFECTIVE   DISEASES. 

spleen,  closely  resembling  miliary  tubercles.  Lomer  recommends 
inoculation  of  guinea-pigs  as  the  most  reliable  method  of  diagnosis. 

Sub-cultures  of  the  bacillus  rapidly  lose  their  virulence.  The 
toxic  products  have  been  already  described  (p.  48).  Mallein  can 
be  prepared  from  a  culture  on  sterilised  potato  by  extracting 
with  glycerine  and  water,  or  from  a  culture  of  the  bacillus  in 
broth.  A  virulent  culture  is  obtained  from  a  glandered  horse, 
or  from  a  guinea-pig  inoculated  with  fresh  virus.  Sub-cultures  are 
prepared  in  glycerine  broth,  and  incubated  at  37°  C.  for  a  month 
or  six  weeks.  If  the  cultures  are  found  to  be  pure  they  are  sterilised 
in  the  usual. way,  in  the  steam  steriliser,  and  by  filtering  through 
porcelain  a  pale,  amber- coloured  liquid  is  obtained.  To  test  for 
glanders,  a  few  drops  (2^  cc.)  are  injected  underneath  the  skin,  in  the 
middle  of  the  side  of  the  neck.  In  healthy  horses  there  is  no  re- 
action, or  a  very  slight  elevation  of  temperature.  In  glandered  horses 
there  is  marked  rise  of  temperature  (101°  to  105°),  considerable  local 
swelling  at  the  seat  of  inoculation,  and  signs  of  general  disturbance, 
while  the  glandered  tumours  become  more  swollen  and  painful.  The 
temperature  of  the  horse  to  be  injected  should  be  taken  night  and 
morning  for  two  or  three  days  before  the  operation ;  and  in  horses 
suffering  from  febrile  disturbance  the  test  should  be  delayed. 
Thomassen  made  a  number  of  experiments  on  horses  suffering  from 
pleurisy,  bronchial  catarrh,  strangles,  and  other  diseases,  without 
any  reaction,  except  in  a  glandered  horse,  used  as  a  control  experi- 
ment. Hunting  and  M'Fadyean,  in  this  country,  have  made  most 
careful  observations  and  experiments,  and  there  is  no  doubt  that 
rnallein  is  a  very  valuable  aid  in  the  diagnosis  of  glanders.  In. 
many  cases  the  reaction  has  been  obtained  in  horses,  and  the 
existence  of  glanders  has  been  discovered  only  after  a  most  searching 
post-mortem  examination.  With  this  means  of  diagnosis  it  is  now 
possible  to  determine  exactly  which  are  the  infected  animals  in  a 
stable  where  the  disease  has  broken  out.  The  animals  can  then 
be  slaughtered,  and  the  disease  prevented  from  spreading. 

Stamping-out  System. — Whatever  may  be  the  stage  of  the 
disease  or  the  extent  or  variety  of  it,  isolation  ought  to  be  carried 
out  in  its  most  complete  form — namely,  slaughter.  The  disease 
might  be  completely  stamped  out,  if  it  were  not  for  the  difficult 
question  of  compensation.  It  can  undoubtedly  be  checked  by  the 
existing  laws. 

In  1869  glanders  was  included  in  the  list  of  contagious  diseases, 
and  the  provisions  with  regard  to  giving  notice  of  the-  disease,  the 
regulation  of  movement,  or  exposure,  and  disinfection,  were  applied  to 


GLANDERS.  455 

horses  suffering  from  glanders.  Under  an  earlier  Act  it  was  made 
an  offence  to  expose  glandered  horses  in  markets  or  on  commons. 
'In  1878  power  to  slaughter  was  incorporated  in  the  Animals  Order. 

(1)  Where  a  person  having  a  horse,  ass,  or  mule  in  his  possession,  or 
under  his  charge,  gives  notice  to  a  constable  that  the  horse,  ass,  or  mule  is 
affected  with  glanders,  or  any  person  is  convicted  of  an  offence  against 
the  Act  of  1878  by  reason  of  his  having  failed  to  give  such  a  notice,  then, 
if  at  any  time  thereafter  it  appears  to  the  Local  Authority,  on  a  special 
report  of  a  Veterinary  Inspector,  that  the  horse,  ass,  or  mule  is  affected 
with  glanders,  and  the  horse,  ass,  or  mule  is  alive  at  the  end  of  fourteen  days 
after  the  receipt  by  the  Local  Authority  of  that  special  report,  the  Local 
Authority  may  serve  on  the  owner  of  the  horse,  ass,  or  mule  a  notice  in 
writing  requiring  him  to  slaughter  it,  or  to  permit  them  to  slaughter  it, 
within  a  time  specified  in  the  notice. 

(2)  If  in  any  case  the  owner  fails  to  comply  with  the  requisition  of 
the  notice  of  the  Local  Authority,  he  shall  be  deemed  guilty  of  an  offence 
against  the  Act  of  1878,  unless  he  shows  to  the  satisfaction  of  the  court 
of  summary  jurisdiction  before  which  he  is  charged  that  the  horse,  ass, 
or  mule,  is  not  affected  with  glanders,  or  that  the  slaughter  thereof  is 
for  any  reason  unnecessary  or  inexpedient. 

(3)  The  provisions  of  this  Article  may  be  put  in  force,  from  time  to 
time,  as  often  as  occasion  requires,  in  relation  to  the  same  horse,  ass,  or 
mule,  on  a  further  special  report  as  aforesaid. 

In  the  order  of  1892  it  was  provided  that  glanders  should  include 
farcy,  and  power  was  given  to  compel  slaughter,  and  to  compensate 
by  payment  of  half  the  value  of  a  diseased  animal,  not  exceeding  £20, 
and  full  value  in  the  case  of  healthy  animals.  Owing  to  objections 
urged  against  the  payment  of  compensation,  another  order  was 
passed,  which  came  into  operation  atgthe  end  of  1894;  the  order  to 
slaughter  being  amended  as  under  : — 

(1)  A  Local  Authority  may  if  they  think  fit  cause  to  be  slaughtered 
any  diseased  horse,  ass,  or  mule,  provided  that  if  the  owner  of  the  horser 
ass,   or  mule  gives  notice  in  writing  to  the  Local  Authority,  or  their 
inspector  or  other  officer,  that  he  objects  to  the  horse,  ass,  or  mule  being 
slaughtered,  it  shall  not  be  lawful  for  the  Local  Authority  to  cause  that 
horse,  ass,  or  mule  to  be  slaughtered  except  with  the  further  special 
authority  of  the  Board  of  Agriculture  first  obtained. 

(2)  A  Local  Authority  may,  if  they  think  fit,  cause  to  be  slaughtered 
any  suspected  horse,  ass,  or  mule,  having  previously  obtained  the  consent 
of  the  owner  thereof. 

(3)  The  Local  Authority  shall  out  of  the  local  rate  pay  compensation 
as  follows  for  any  horse,  ass,  or  mule  slaughtered  under  this  article  : — 

(a)  Where  the  horse,  ass,  or  mule  was  diseased  the  compensation  shall 


456  INFECTIVE   DISEASES. 

be  such  sum  as  the  Local  Authority  think  expedient,  being  a 
minimum  in  the  case  of  a  horse  of  two  pounds,  and  in  the  case  of 
an  ass  or  mule  of  ten  shillings  ;  provided  that  in  no  case  shall  the 
amount  of  compensation,  if  above  the  said  minimum,  exceed  one- 
fourth  of  the  value  of  the  animal  immediately  before  it  became 
diseased. 

(b)  In  every  other  case  the  compensation  shall  be  the  value  of  the 
horse,  ass,  or  mule  immediately  before  it  was  slaughtered. 


CHAPTER  XXXII. 

TETANUS,    RABIES,    AND   LOUPING-ILL. 

TETANUS. 

TETANUS  is  a  communicable  disease  of  man  and  the  lower  animals, 
characterised  by  spasmodic  contraction  of  the  muscles.  It  is  com- 
monly the  result  of  .an  injury,  and  occurs  especially  after  wounds 
produced  by  means  of  splinters  of  wood  or  contaminated  with  earth 
or  dust,  and  may  follow  after  surgical  operations. 

Carle  and  Rattone  first  showed,  in  1884,  that  the  disease  could 
be  communicated  from  man  to  other  animals.  Rabbits  inoculated 
with  pus  from  a  case  in  man  developed  tetanus,  and  from  these 
rabbits  the  disease  was  conveyed  to  others.  Nicola ier,  the  following 
year,  found  that  mice  and  rabbits  inoculated  with  earth  often 
contracted  tetanus,  and  that  the  pus  which  formed  at  the  seat  of 
inoculation  contained,  amongst  other  organisms,  characteristic  bacilli. 
Pure  cultures  were  first  obtained  by  Kitasato. 

Bacillus  of  Tetanus. — Slender,  straight  rods,  and  filamentous 
forms.  Spore  formation  takes  place  at  the  end  of  a  bacillus,  giving 
it  a  drumstick  appearance.  They  stain  with  aniline  dyes,  but  best 
with  Neelsen's  solution,  or  by  Gram's  method.  By  the  Ziehl-Neelsen 
method  and  methylene-blue,  the  spores  can  be  stained  red,  in 
contrast  to  the  bacilli,  which  are  stained  blue.  The  bacilli  are 
anaerobic,  liquefy  gelatine,  and  are  slightly  motile.  They  can  be 
grown  at  the  ordinary  temperature,  but  most  readily  at  the  tem- 
perature of  the  blood,  in  an  atmosphere  of  hydrogen,  especially  after 
the  addition  of  1  or  2  per  cent,  of  grape  sugar  to  the  nutrient 
medium.  The  young  colonies  on  plate-cultivations  somewhat 
resemble  those  of  Bacillus  subtilis.  They  have  an  opaque  centre, 
and  are  surrounded  by  fine  rays,  extending  in  all  directions  like 
thistle-down.  In  the  depth  of  gelatine  a  ray-like  growth  occurs 
in  the  lower  part  of  the  track  of  the  needle.  The  gelatine  is  lique- 
fied very  slowly,  and  gas  is  given  off.  The  cultures  possess  a 

457 


458 


INFECTIVE    DISEASES. 


characteristic  odour.  In  slightly  alkaline  broth,  and  peptone  with 
alkaline  reaction,  in  an  atmosphere  of  hydrogen,  the  gas  formed 
will  be  sufficient  to  break  the  flask  if  it  is  sealed  up.  Kitasato 
obtained  his  cultures  from  pus,  by  taking  advantage  of  the  resist- 
ance of  the  spores  to  high  temperatures.  By  raising  cultures 
to  80°  C.  for  three-quarters  of  an  hour,  the  micrococci  and  bacilli 
in  the  mixed  culture  were  destroyed,  while  the  spores  of  the 
tetanus  bacillus  retained  their  vitality,  and  then  sub-cultures  were 
obtained  in  a  pure  state.  The  spores  are  said  to  be  killed  by 
exposure  to  steam  for  five  minutes.  A  5  per 
cent,  solution  of  carbolic  acid  with  '5  per  cent, 
of  hydrochloric,  will  destroy  the  spores  in  two 
hours.  Kitasato  and  Weyl  obtained  tetanin 
!l  from  pure  cultures  of  the  bacillus,  Brieger 
having  previously  obtained  it  from  impure 
cultures.  A  tetano-toxin,  indol  and  phenol,  and 
butyric  acid  are  also  found.  Brieger  and 
Frankel  attribute  the  pathogenic  properties  to 
a  tox-albumin.  These  products  have  been  de- 
scribed more  fully  in  a  previous  chapter  (p.  41). 
A  pure-culture  produces  tetanus  in  a  mouse 
in  twenty-four  hours,  and  rabbits,  guinea-pigs, 
and  rats  can  also  be  infected.  No  pus  forms 
at  the  seat  of  inoculation,  as  after  inoculation 
of  earth,  but  the  spasms  commence  in  the 
muscles  nearest  to  the  seat  of  inoculation.  A 
trace  of  a  broth  culture  will  kill  a  guinea-pig, 
the  symptoms  developing  in  three  days. 

Kitasato  succeeded  in  making  animals  im- 
mune to  tetanus,  and  subsequently  the  discovery 
was  made  that  the  blood  in  immune  animals 
FIG.  186.— PURE-CUL-  w^  produce  immunity  in  other  animals,  the 
TURK  OF  TETANUS  explanation  being  that  the  toxic  principle  of 
BACILLI  IN  GRAPE-  the  tetanus  bacillus  induces  the  formation  of 
SUGAR  GELATINE. 
Four  days  old.  tetanus  antitoxin ;  and  it  equal  parts  ot  the 

(FRANKEL     AND   serum  of  an  immune  animal,  and  a  fatal  dose  of 
tetano-toxin,  are  together  injected  in  to  a  healthy 

guinea-pig,  tetanus  will  not  follow,  showing  that  the  virus  has 
been  neutralised.  Tizzoni  and  Cattani  found  that  blood  from  an 
immunised  dog  was  not  only  capable  of  completely  neutralising  the 
toxic  power  of  filtered  cultures,  but  that  the  injection  of  the  blood- 
serum  produced  immunity  in  otherwise  susceptible  animals,  except 


DESCRIPTION    OF    PLATE    XXII. 
Bacillus  tetani. 

FIG.  1.— From  a  cover-glass  preparation  of  a  pure-cultivation  of  the  tetanus 
bacillus  in  broth ;  stained  with  Neelsen's  carbolised  fuchsine.  x  1200. 
Lamplight  illumination, 

FIG.  2.— From  a  cover-glass  preparation  from  the  same  source ;  stained  with 
Neelsen's  solution  and  methylene  blue,  x  1200.  Lamplight  illumination. 


A 


BACILLUS   TETANI 


DESCRIPTION    <»K    PLATE    XXII. 

Bacillus  tetani. 


FIG.  1.  .From  ;i  mvrr-^lass  preparation  of  a  purr-cultivation  of  the  tetanus 
bacillus  in  broth;  stained  with  K  ,  >ed  fuchsine.  s.  1200. 

Lamplight  illumination. 

Fit;.  2.  —  From  a  cover-glass  preparation  from  the  sam«  source  ;  stained  with 
-en's  solution  and  methylene  blue,  x  1200.  Lamplight  illumination. 


Plate 


A 


2. 


BACILLUS   TETANI 


Brvolu.Day  &Son,UlK. 


460  INFECTIVE   DISEASES. 

several  months,  and  in  rare  cases  a  much  longer  period.  The  wound 
from  a  bite  may  have  healed,  and  may  again  become  inflamed, 
and  symptoms  follow,  owing  to  the  poison  aft'ecting  the  brain,  spinal 
cord,  or  the  peripheral  nerves.  In  the  dog  the  disease  appears  in 
two  forms —raging  madness  and  dumb  madness ;  and  the  identity  of 
the  virus  in  the  dog  and  in  man  is  shown  by  the  fact  that  virus 
from  man  can  produce  both  forms  of  the  disease  in  the  dog.  The 
virus  may  be  obtained  by  inoculation  of  the  saliva  of  a  rabid  dog ; 
but  this  method  is  uncertain,  as  other  micro-organisms  are  present. 
Pasteur  endeavoured  to  obtain  it  in  a  pure  state,  and  was  able 
to  demonstrate  that  the  spinal  cord,  the  brain,  and  the  nerves 
contain  the  virus.  It  was  also  found  that  by  direct  inoculation  of 
the  nervous  system  the  most  certain  results  followed. 

Bacteria  in  Rabies. — Cocci  have  been  described  in  connection 
with  hydrophobia  by  Fol,  Babes,  and  Dowdeswell.  The  cocci,  it  is 
said,  were  observed  in  sections  of  the  spinal  cord  of  rabid  dogs.  The 
descriptions  given  by  different  observers  vary  considerably,  and  there 
is  not  any  particular  coccus  constantly  associated  with  the  disease. 
Nor  have  any  of  the  bacteria  been  isolated  from  the  diseased  animal, 
which  were  alleged  to  be  present  in  stained  preparations.  By  many, 
hydrophobia  is  believed  to  be  due  to  the  presence  of  a  micro- 
organism, but  at  present  the  nature  of  the  contagium  is  unknown. 

Protective  Inoculation. — -Pasteur  found  that  a  dog  inoculated 
under  the  dura- mater  with  virus  from  the  spinal  cord  of  a  rabid  animal 
will  develop  rabies,  as  a  rule,  within  eighteen  days.  By  trephining 
rabbits  and  inoculating  the  virus,  and  by,  in  the  same  way, 
transmitting  the  virus  from  rabbit  to  rabbit,  the  incubation  period 
gradually  shortens,  until  it  is  reduced  to  six  or  seven  days.  The  virus 
has  then  reached  its  maximum  virulence  in  the  rabbit,  and  is  "  fixed." 
Pasteur  then  studied  the  possibility  of  producing  immunity.  The 
medulla  of  a  rabbit,  containing  the  virulent  virus,  was  suspended  in 
a,  glass  bottle  over  caustic  potash  at  a  temperature  of  25°  C.  If  a 
number  of  spinal  cords  were  thus  treated,  and  examined  from  day 
to  day,  it  was  found  that  they  gradually  lost  their  virulence, 
becoming  completely  inert  in  from  sixteen  to  twenty  days.  A  series 
of  cords  was  thus  obtained  with  diminishing  virulence  ;  by  injecting 
subcutaneously  an  infusion  of  rabid  spinal  cord  crushed  in  broth, 
and  beginning  with  an  inert  cord  on  the  first  day  and  using  the 
next  in  the  series  on  the  second  day,  and  so  on  till  a  fresh  spinal 
cord  could  be  injected,  it  was  found  that  dogs  were  rendered 
insusceptible  to  the  strongest  virus,  administered  by  inoculation  or 
by  exposing  them  to  the  bites  of  rabid  dogs.  Dogs  have  usually 


RABIES.  461 

an  incubation  period  of  several  weeks,  and  Pasteur  conceived  that  it 
would  be  possible  to  anticipate  the  symptoms,  which  would  naturally 
follow  in  a  dog  which  had  been  bitten  or  inoculated,  by  giving 
them  a  mild  form  of  hydrophobia  by  the  injection  of  attenuated 
virus  of  short  incubation  period.  These  experiments  showed  that 
it  was  possible  to  do  this,  and  the  outcome  was  the  introduction  of 
a  system  of  protective  inoculation  in  the  human  subject.  Pasteur 
succeeded  in  giving  immunity  from  hydrophobia  to  about  fifty  dogs 
of  every  age  and  breed. 

In  1885  Joseph  Meister,  a  boy  nine  years  of  age,  bitten  badly 
by  a  mad  dog  upon  the  hands,  legs,  and  thighs,  was  brought  to 
Pasteur.  At  a  post-mortem  examination  of  the  dog,  its  stomach 
was  found  full  of  bits  of  hay,  straw,  and  wood,  and  it  had  been 
unquestionably  rabid.  On  July  6th,  sixty  hours  after  Meister  had 
been  bitten,  a  syringe  full  of  marrow  from  a  rabbit  which  had  died 
on  June  21st,  and  therefore  fifteen  days  old,  was  injected  beneath 
the  skin  over  the  right  hypochondriac  region.  The  next  morning 
Meister  was  inoculated  with  a  spinal  cord  fourteen  days  old,  and  so 
on  every  day,  till  on  the  sixteenth  a  cord  only  one  day  old  was  used. 
So  many  injections,  however,  need  not  have  been  given,  as  it  was 
subsequently  found  that  the  spinal  marrows  injected  during  the 
first  five  days  were  inert  when  tested  on  rabbits.  The  marrows  of 
the  next  five  days  showed  an  ascending  scale  of  virulency,  until,  on 
the  last  two  days  of  the  treatment,  Meister  had  been  inoculated 
with  a  virus  so  virulent  that  it  was  capable  of  causing  hydrophobia 
in  dogs  after  ten  days'  incubation.  Meister  remained  completely 
free  from  hydrophobia.  From  that  time  to  the  present  day  many 
thousands  of  patients  have  been  treated  in  Paris  by  slightly  modified 
methods,  and  it  is  very  generally  believed  that  a  real  prophylactic 
agent  has  been  discovered. 

Pasteur  suggested  that  the  rabic  virus  might  consist  of  two 
distinct  substances — a  living  virus  capable  of  developing  in  the 
nervous  system,  and  a  secondary  product  which,  in  sufficient  pro- 
portions, might  have  the  property  of  hindering  the  development 
of  the  living  virus.  The  nature  of  this  living  virus  is  quite 
unknown. 

According  to  a  return  of  the  inoculations  at  the  Pasteur  In- 
stitute, the  total  number  of  persons  treated  in  1895  was  1,523,  of 
whom  five  died.  In  three  cases  the  symptoms  of  hydrophobia 
occurred  within  a  fortnight  of  the  last  inoculation.  If  these  three 
cases  are  omitted,  the  number  of  persons  treated  is  reduced  to  1,520 
and  the  deaths  to  two.  The  results  are  shown  in  the  following  table,. 


462 


INFECTIVE   DISEASES. 


for  the  nine  years  previous  to  1895,  during  which  Pasteur's  method 
has  been  in  operation  : — 


Year. 

Number  of 
persons 
inoculated. 

Number  of 
deaths. 

Rate  of 
Mortality. 

1886     

2,671 

25 

0-94 

1887     

1,770 

14 

0-79 

1888     

1,622 

9 

0-55 

1889     

1,830 

7 

0-38 

1890     

1,540 

5 

0-32 

1891     

1,559 

4 

0-25 

1892     ... 

1,790 

4 

0-22 

1893     

1,648 

6 

0-36 

1894     ... 

1,387 

7 

0-50 

1895     ... 

1,520 

2 

0-13 

Of  the  1,520  persons  treated,  156  were  bitten  on  the  head  or 
face,  829  upon  the  hands,  and  535  on  other  parts  of  the  body  ;  122 
were  bitten  by  animals  experimentally  proved  to  be  rabid,  949  by 
animals  declared  by  veterinary  certificate  to  be  rabid,  and  449  by 
animals  supposed  to  be  rabid. 

Babes  at  Bucharest  inoculated  300  persons  in  one  year,  and 
claimed  to  have  reduced  the  mortality  to  *4  per  cent. 

Stamping-out  System. — There  is  every  reason  to  believe  that 
rabies  could  be  sta raped  out  in  England  in  six  months,  if  a  general 
order  for  muzzling  were  enforced,  and  all  ownerless  dogs  were 
slaughtered.  It  is  the  ownerless  cur,  the  vagrant  dog,  which  is 
mainly  responsible  for  the  spread  of  rabies ;  and  if  a  general  muzzling 
order  cannot  be  put  into  force,  it  would  undoubtedly  check  the  disease 
if  all  dogs  were  compelled  to  wear  a  collar  with  the  name  and  address 
of  the  owner,  and  all  dogs  without  owners  were  destroyed. 


LOUPING-ILL. 

Louping-ill  is  regarded  by  some  as  an  infective  disease.  It  is 
a  disease  of  sheep,  characterised  by  symptoms  due  to  an  affection  of 
the  central  nervous  system.  The  symptoms  consist  in  contractions 


LOU  PING-ILL.  463 

of  the  muscles  of  the  head  and  limbs,  loss  of  co-ordination  and  finally 
complete  loss  of  the  power  of  movement.  The  name  is  derived  from 
the  peculiar  jumping  movements  in  the  early  stage. 

Klein  and  M'Fadyean  independently  investigated  this  disease. 
Klein  found  bacteria  in  the  cerebral  fluid.  No  micro-organisms 
were  found  in  the  blood.  Special  attention  was  drawn  to  a 
bacterium  which  was  found  by  Klein  in  six  out  of  seventeen  cases, 
and  to  a  micrococcus  by  M'Fadyean. 

Bacteria  in  Louping-Ill — Klein's  Bacterium.  Oval  cocci  and 
rods  *6  to  1  /A  in  length,  *2  to  '3  /A  in  breadth.  Colonies  in  gelatine, 
yellowish  by  reflected  light,  are  brown  by  transmitted  light.  On  the 
surface  of  gelatine  the  bacteria  form  a  film,  which  is  crenated  at 
the  edge,  and  thick  in  the  middle,  at  first  grey  and  later  yellowish. 
In  the  depth  of  gelatine  a  filament  forms,  composed  of  closely 
aggregated  minute  greyish  colonies,  and  a  prominent  yellow  growth 
occurs  on  the  free  surface.  The  gelatine  is  not  liquefied.  The 
bacteria  grow  in  milk,  and  broth  becomes  turbid  in  two  days,  and 
there  is  a  copious  flocculent  greyish  precipitate. 

Injection  of  broth  cultures  subcutaneously  in  rabbits,  guinea- 
pigs,  and  mice,  produced  no  result,  except  local  swelling  at  the  seat 
of  inoculation,  which  subsided  without  causing  any  constitutional 
.symptoms.  The  results  were  equally  negative  when  the  cultures 
were  injected  subcutaneously  in  lambs. 

M'Fadyean' a  Micrococcus. — Cocci  '3  //,  in  diameter.  The  colonies 
are  flat,  nearly  circular,  and  have  a  smooth  edge.  In  old  colonies 
the  centre  appears  as  a  dark  spot.  Gelatine  is  rapidly  liquefied, 
and  a  nearly  colourless  precipitate  forms  at  the  bottom  of  the 
tube.  Cultures  on  the  surface  of  agar  have  a  faint  yellow  tinge. 
On  potato  the  colour  is  deeper  but  the  growth  not  so  well  marked. 
Milk  is  coagulated.  In  broth  there  is  an  abundant  growth  render- 
ing the  liquid  turbid  and  depositing  a  white  precipitate.  The  micro- 
cocci  stain  by  Gram's  method.  Inoculated  in  rabbits  or  guinea-pigs, 
they  produce  suppuration ;  in  horses  and  bovines,  an  inflammatory 
swelling  results  without  suppuration.  They  produce  abscesses  in 
sheep  and  lambs.  The  cocci  were  isolated  from  abscesses  in  lambs 
suffering  from  louping-ill.  Though  it  is  admitted  that  louping-ill 
belongs  to  the  class  of  infective  diseases,  there  can  be  no  doubt  from 
these  experiments  that  the  nature  of  the  contagium  is  unknown. 


CHAPTER   XXXIII. 

FOOT-ROT. 

SHEEP  are  subject  to  several  diseases  which  are  classed  as  foot -rot. 
There  is  one  form,  known  as  contagious  foot-rot,  which  prevails  in 
certain  localities,  especially  on  wet  land.  Brown  describes  the 
disease  as  primarily  a  disease  of  the  skin,  inducing  exfoliation  of 
the  cuticle,  and  exudation  of  fluid  containing  epithelial  scales.  The 
inflammation  extends  to  the  membrane  of  the  foot,  leading  to  exfolia- 
tion of  the  hoof,  and  development  of  epithelial  scales,  which  form  an 
imperfect  horny  layer  on  the  diseased  membrane.  In  one  outbreak 
investigated  by  Brown,  the  disease  in  the  early  stage  was  confined 
to  the  skin  between  the  digits  of  the  fore-feet;  the  surface  was 
red,  tumid  and  pulpy,  and  white  purulent  matter  existed  on  the 
inflamed  parts.  Later,  the  hoof  grew  to  an  extraordinary  length, 
fungoid  growths  made  their  appearance,  developing  into  foot-rot  in 
an  advanced  form. 

In  France,  according  to  Fleming,  the  contagious  character  of 
this  disease  has  long  been  recognised.  So  long  ago  as  1805  Pictet 
imported  200  half-bred  merino  sheep,  some  of  which  were  infected 
with  foot-rot,  and  placed  them  with  200  healthy  sheep,  and  in  a 
short  time  all  the  sheep  were  infected.  Several  years  later  Favre 
and  Sorillon  carried  out  investigations  which  conclusively  proved 
the  infective  nature  of  the  virus.  Among  other  experiments  it 
was  found  that  when  healthy  sheep  wrere  inoculated  in  the  feet 
with  virus  from  diseased  sheep,  the  disease  was  communicated. 

Contagious  foot-root  may  be  spread  by  healthy  sheep  receiving 
the  virus  from  infected  sheep  in  fairs  and  markets.  Ships,  railway- 
trucks,  and  carts  in  which  diseased  sheep  have  been  conveyed,  unless 
subsequently  thoroughly  disinfected,  may  be  the  means  of  trans- 
mitting the  virus  to  healthy  sheep.  Healthy  sheep  turned  into 
pastures  quite  recently  occupied  by  diseased  sheep  may  be  inoculated 
from  the  discharges  from  the  feet  of  the  diseased  sheep,  which 
contaminate  the  grass  and  soil. 

464 


FOOT-ROT. 


465 


Law  believed  that  the  disease  arose  from  an  undiscovered  micro- 
organism, which  was  probably  present  in  infected  pastures.  Others 
in  this  country  have  disputed  the  contagious  character  of  the  disease, 
and  considered  that  the  same  conditions  of  the  pasture  which 
produced  the  disease  in  a  flock  would  produce  it  again  in  imported 
animals,  which  would  account  for  the  apparent  contagiousness. 

Brown,  in  order  to  test  the  question  of  contagion,  placed  infected 
sheep  in  a  pen,  the  bottom  of  which  was  covered  with  straw  which 
was  not  removed  while  the  experiments  were  in  process.  Three 
healthy  sheep,  from  a  locality  where  foot-rot  was  unknown,  were 
placed  with  the  infected  sheep.  At  the  end  of  ten  days  the  feet 
of  the  sound  sheep  were  still  healthy.  Subsequently  two  of  the 
sheep  were  inoculated,  and  it  was  found  that  the  virus  introduced 


FIG.  187.— FOOT  OP  SHEEP  SHOWING 
DISEASE  OF  HORN  (BROWN). 


FIG.  188.—  SECTION  THROUGH  THE 
FOOT  SHOWING  A  CRACK  EX- 
TENDING THROUGH  THE  WALL. 


subcutaneously  in  the  vicinity  of  the  foot  produced  the  incipient 
stage  of  the  disease.  On  making  further  experiments  the  contagious 
nature  of  one  form  of  foot-rot  was  established,  but  it  appears  that 
the  contagious  property  is  only  developed  after  a  long  period  of 
exposure,  and  under  certain  conditions.  On  a  dry  soil  the  disease 
will  quickly  subside,  but  on  moist  land  the  contagious  form  of  foot- 
rot  may  be  communicated  by  simple  contact,  in  from  six  weeks  to 
three  months. 

From  these  and  other  experiments  Brown  has  drawn  the 
following  conclusions : — 

1.  That  so  far  as  the  evidence  goes  it  justifies  the  statement  that 
foot-rot  is  a  contagious  disease ;  the  infective  matter  being  active 
when  brought  into  contact  with  the  skin  between  the  claws,  or 

30 


466 


INFECTIVE   DISEASES. 


if 
II 


T 


OF  THE 

••riVERSlTY 


FOOT-ROT. 


467 


when  introduced  into  the  system  by  inoculation,  and  probably  when 
taken  in  by  the  mouth  from  contaminated  pastures. 

2.  That  it  cannot  be  produced  by  long-continued  exposure   to 
imdrained,  moist  soils,  with  an  abundant,  coarse  and  wet  herbage. 

3.  That  animals  exposed  to  these  conditions  for  many  months, 
.and  resisting  entirely  the  influences  named  above,  contract  foot-rot 


FIG.  191.—  DISTORTION  OF  HOOF  IN  AN  ADVANCED  FORM  OF  FOOT-ROT  (BROWN). 

in  from  fourteen  to  twenty-one  days  on  being  placed  among  sheep 
suffering  from  the  disease. 

4.  That  sheep  affected  with  foot -rot  may  improve,  and  from 
time  to  time  become  worse ;  and  finally  may  recover  and  present  a 
perfectly  healthy  condition  of  foot,  notwithstanding  that  they  have 
been  kept  the  whole  period  under  the  conditions  which  induced  the 
disease. 


468  INFECTIVE   DISEASES. 

5.  That  the  contagium  of  foot-rot  remains  for  some  time  in 
the  system  (ten  to  twenty  days  and  longer)  without  any  indication 
of  disease  appearing  in  the  skin  between  the  claws.  An  infected 
sheep  may  therefore  escape  detection  even  by  an  expert,  and  may 
introduce  foot-rot  into  a  sound  flock. 

One  attack  does  not  confer  immunity.  The  disease  has  been 
known  to  recur  in  sheep  which  have  only  recently  recovered  from 
an  outbreak. 

The  disease  is  no',  communicable  to  other  animals,  including 
man.  The  flesh  is  harmless,  but  as  in  severe  cases  the  sheep  are 
emaciated,  the  carcass  is  in  consequence  of  little,  if  of  any,  value 
as  food. 

The  nature  of  the  contagium  is  unknown. 

Stamping-out  System. — Sheep  should  not  be  allowed  to  be 
moved  from  an  infected  district  except  for  slaughter.  When  sheep 
are  purchased  to  add  to  the  stock  on  a  farm,  they  should  be 
isolated  for  two  or  three  weeks,  and  carefully  watched  before  they 
are  allowed  to  mix  with  other  sheep.  If  this  disease  is  detected 
in  a  flock,  every  animal  should  be  carefully  examined,  and  any 
suspicious  as  well  as  any  diseased  sheep  should  be  completely  isolated 
from  the  rest.  Fleming  recommends  that  those  which  have  been 
in  contact,  though  still  apparently  quite  healthy,  should  as  a  pre- 
cautionary measure,  be  made  to  pass  through  a  trough  containing 
a  solution  of  chloride  of  lime  to  a  'depth  of  about  four  inches. 
The  solution  is  made  by  adding  one  or  two  pounds  of  chloride  of 
lime  to  two  buckets  of  rain-water.  If  the  trough  is  placed  at  the 
entrance  to  the  sheepfold,  the  sheep  will  be  compelled  to  traverse 
it  at  least  twice  a  day.  It  is  also  recommended,  when  diseased 
sheep  are  treated  by  this  plan,  that  after  recovery,  all  manure 
in  the  fold  should  be  removed  and  destroyed,  and  the  soil  dug  up 
to  the  depth  of  six  inches,  or  lime  freely  spread  over  the  surface. 
Troughs  and  hurdles  must  be  thoroughly  disinfected,  and  buildings 
freely  ventilated  after  similar  treatment.  No  locality  can  be  con- 
sidered free  from  suspicion  until  one  or  two  months  have  elapsed 
since  the  recovery  of  the  last  case 


CHAPTEE   XXXIV. 

FOUL-BROOD—INFECTIOUS  DISEASE   OF   BEES  IN   ITALY — PEBRINE — 
FLACHERIE — INFECTIOUS    DISEASE   OF   CATERPILLARS. 

FOUL-BROOD  IN  BEES. 

FOUL-BROOD  is  a  contagious  disease  attacking  bees  and  especially  the 
larvae.  The  larvae  rapidly  lose  their  healthy  appearance,  die  and 
decompose,  turning  into  a  coffee-coloured  mass.  The  cells  of  the 
honeycomb  are  mapped  out  by  the  dark- brown  cappings  of  the  cells 


FIG.  192.— DISEASED  CQMB  (COWAN). 

containing  the  diseased  larvse.  The  decomposition  is  associated  with 
the  production  of  an  unpleasant  odour,  which  can  be  detected  at 
some  distance  from  an  infected  hive.  The  disease  has  been  known 
from  very  early  times.  Preuss  investigated  it  microscopically,  and 
attributed  it  to  "  micrococci."  These  were  really  the  spores  of  a 
bacillus,  which  was  first  observed  by  Cohn.  Later,  Cheshire  and 

469 


470 


INFECTIVE    DISEASES. 


FIG.  193.— SPORES  OF  BACILLUS 
ALVEI. 


Cheyne   investigated   foul-brood,    isolated   the    bacillus,    and   fully 

described  its   morphological  and  biological  characters   in   nutrient 

media.  By  removing  the  cap  of 
one  of  the  diseased  cells,  the  de- 
composing larva  can  be  withdrawn, 
and  cover-glass  preparations  will 
reveal  delicate  bacilli  and  large  oval 
spores.  The  bacilli  can  be  readily 
isolated  and  cultivated  in  nutrient 
gelatine. 

Bacillus  alvei  (Cheshire  and 
Cheyne).  Hods  varying  in  size, 
and  forming  large  oval  spores. 
They  are  motile  and  possess  flagella. 
Cultivated  in  nutrient  gelatine  in 

test-tubes,  a  delicate   ramifying  growth   appears    on    the   surface, 

and  irregular  whitish  masses  arise  along   the  needle  track.     Pro- 
cesses shoot  out  from  these  masses,  and 

extend   through    the    gelatine    for    long 

distances.     They  are  thickened  at  points 

in  their  course,  and  are  clubbed  at  the 

ends.     The  gelatine  is  gradually  liquefied, 

and  the  bacilli  form  a  loose,  white,  floccu- 

lent  deposit  at  the  bottom  of  the  tube. 

The  liquid  in  the  tube  becomes  yellowish 

in  colour  after  a  time,  and  gives  off  an 

odour  of  stale,  but  not  ammoniacal,  urine. 

On  the   surface  of  nutrient  gelatine  the 

bacilli  grow  out  in  chains  of  rods  in  single 

file,   or  of   rows  of   several  side  by  side. 

The  processes  which  are  formed  tend  to 

curve,  and  at  a  short  distance  from  the 

track  of  the  needle  form  a  distinct  circle, 

from   which  another  process   grows  out, 

and   a    fresh    circle    is    developed.     The 

gelatine   in   the   vicinity   of     the    bacilli 

gradually    liquefies,     and     channels    are 

formed    in    the    gelatine    in    which  the 

bacilli   move    backwards    and    forwards. 

On   nutrient  agar-agar  a    whitish    layer 

develops,    consisting   of    bacilli    arranged  side    by  side,  which  in  a 

few  days  are  replaced  by  rows  of  spores  similarly   arranged. 


FIG  194.— PURE-CULTURE  IN 
NUTRIENT  GELATINE,  x  4 
(CHESHIRE  AND  CHEYNE). 


On 


INFECTIOUS   DISEASE   OF   BEES   IN  ITALY.  471 

potatoes  they  form  a  dryish,  yellow  layer,  and  in  milk  a  tremulous 
jelly.  A  cultivation  of  the  bacillus  in  milk,  sprayed  over  a  honey- 
comb containing  a  healthy  brood  of  bee  larvae,  produced  foul-brood. 
Adult  bees  fed  on  material  containing  bacilli  became  infected. 
Inoculation  of  mice  and  rabbits  with  the  bacillus  gave  doubtful 
results. 

Stamping-out  System.— The  infected  bees,  combs,  frames  and 
quilts  must  be  destroyed,  and  the  hives  thoroughly  disirifected,  as 
this  is  the  only  way  in  which  the  resistant  spores  can  be  got  rid  of. 
Cowan  believes  that  if  foul-brood  were  under  Government  inspection 
and  infected  hives  were  destroyed,  the  disease  could  be  stamped  out,. 


FIG.  195.—  CULTIVATION  ON  THE  SURFACE  OF  GELATINE,  x  80 
(CHESHIRE  AND  CHEYNE). 


INFECTIOUS  DISEASE  OF  BEES  IN  ITALY. 

In  Italy  bees  are  subject  to  another  infectious  malady,  and 
Canestrini  has  found  bacilli  in  the  bees  and  in  the  larvae,  which  are 
believed  to  be  the  cause  of  the  malady. 

Bacillus  of  Infectious  Disease  of  Bees  (Canestrini). — Hods 
2  /x  in  width  and  4  to  6  /A  in  length,  occurring  singly,  in  pairs 
and  in  chains,  sometimes  capsulated.  They  are  motile,  and  spore - 
formation  is  present.  They  liquefy  gelatine,  colouring  the  liquid 
pink  and  forming  a  white  deposit.  On  agar  they  form  a  white 
growth,  and  on  potato  la  claret-coloured  layer.  Cultures  are  said 
to  be  capable  of  producing  the  disease  in  bees  and  larvae. 

PEBRINE. 

The  silkworm  disease  known  in  France  as  pebrine  is  characterised 
by  the  appearance  of  black  patches  on  the  skin  of  the  worms.  It 
was  investigated  by  Cornalia,  Nageli,  and  Pasteur.  Pasteur's 

~ 
.     RA . 

OF    THK 

UNIVERSITY 


472  INFECTIVE   DISEASES. 

prophylactic    measures    have   been   described    in    another    chapter 

(p  "<)• 

Panhistophyton  ovatum.  (Lebert.  Nosema  bombycis,  Micro- 
coccus  ovatus,  Corpuscles  du  ver-a-soie). — Shining  oval  cocci,  2  to  3  /x 
long,  2  /x  wide,  singly  and  in  pairs,  or  masses ;  or  rods,  2 '5  /x  thick, 
and  twice  as  long.  They  multiply  by  subdivision.  They  were 
experimentally  proved  to  be  the  cause  of  pebrine,  gattine,  maladie 
des  corpuscles  or  Flecksucht ;  and  were  discovered  in  the  organs  of 
diseased  silkworms,  as  well  as  in  the  pupae,  moths,  and  eggs. 

Metchnikoff  believes  that  these  micro-organisms  are  not  bacteria, 
but  psorosperms. 

FLACHERIE. 

Silkworms  are  also  subject  to  a  very  destructive  disease  known 
as  jlacherie,  flaccidezza,  maladie  de  morts  blancs.  The  worms  cease 
feeding,  die  and  become  a  putrid  mass.  The  disease  is  dependent 
upon  bad  hygienic  conditions,  and  is  very  infectious.  The  cause 
has  not  been  determined  with  certainty,  but  it  has  been  attributed 
to  a  streptococcus. 

Streptococcus  bombycis  (Mikrozyma  bombycis,  Bechamp).— 
Oval  cocci  1-5  /x  diarn.,  singly,  in  pairs,  and  in  chains.  They  are 
said  to  be  present  in  dust  from  infected  localities. 

DISEASE  OF  CATERPILLARS. 

Forbes  has  described  an  infectious  disease  of  the  larvae  of  a 
caterpillar  (Picris  rapce).  Cocci  which  were  found  singly  and  in 
masses,  were  regarded  as  the  cause  of  the  malady. 


PAET    III. 

SYSTEMATIC    AND    DESCRIPTIVE. 


473 


CHAPTEK   XXXV. 

CLASSIFICATION   AND   DESCRIPTION    OF    SPECIES. 

IN  reviewing  the  history  of  the  various  classifications  which  have 
from  time  to  time  been  proposed,  we  shall  see  that  the  gradual 
improvements  in  the  means  of  studying  such  minute  objects, 
the  methods  of  cultivating  them  artificially,  and  of  studying  their 
chemistry  and  physiology,  and  the  ever- increasing  revelations  of  the 
microscope,  have  resulted  in  establishing  these  microscopic  objects  as 
members  of  the  vegetable  kingdom,  ranking  among  the  lowest  forms 
of  fungi,  but  with  regard  to  the  division  into  genera  and  species  we 
are  still  in  a  position  of  doubt  and  uncertainty. 

Miiller,  in  1773,  was  the  first  to  suggest  a  classification.  He 
established  two  genera,  Monas  and  Yibrio,  and  grouped  them  with 
the  Infusoria.  In  1824  Bory  de  Saint  Vincent  also  attempted 
a  classification;  but  it  was  not  until  Ehrenberg  in  1838,  and 
Dujardin  in  1841,  worked  at  the  subject,  that  a  scientific  distinction 
of  species  was  attempted. 

Ehrenberg  described  four  genera  : — 

I.  Bacterium     .  .  filaments  straight,  rigid. 

II.  Vibrio  .  .  filaments  snake-like,  flexible. 

III.  Spirillum      .  .  filaments  spiral,  rigid. 

IV.  Spirochaeta  .  .  filaments  spiral,  flexible. 

Djijardin  united  Spirillum  and  Spirochseta,  and  classed  them 
thus  : — 

I.  Bacterium  .  .  filaments  rigid,  vacillating. 

II.  Vibrio          .  .  filaments  flexible,  undulatory. 

III.  Spirillum    .  .  filaments  spiral,  rotatory. 

Bacteria  were  still  considered  as  Infusoria,  but  in  1852  Perty 
maintained  that  some  of  the  smallest  living  organisms  belonged  to- 
the  animal  and  others  to  the  vegetable  kingdom,  and  that  Vibrios 
without  question  belonged  to  the  latter.  In  1853  Robin  pointed  out 

475 


476  SYSTEMATIC. 

the  affinity  of  the  Bacteria  and  Vibrios  to  Leptothrix  ;  and  Davaine, 
in  1859,  insisted  that  the  Vibrios  were  vegetables,  and  were  in 
fact  allied  to  the  Algse. 

Since  that  time  a  flood  of  light  has  poured  in  upon  this  subject 
through  the  writings  of  Hoffmann,  Pasteur,  Cohn,  Rabenhorst, 
Hallier,  Billroth,  Warming,  Nageli,  Magnin,  Marchand,  Sternberg, 
Van  Tieghem,  Koch,  Flugge,  De  Bary,  Zopf,  Biichner,  Hueppe, 
Marshall  Hall,  and  many  others  who  have  studied  the  morphology, 
life-history,  and  classification  of  bacteria. 

Of  all  these  writers  we  are  most  indebted  to  Cohn,  not  only 
on  account  of  his  researches,  which  extended  over  many  years,  but 
also  for  his  system  of  classification,  which  has  since  been  almost 
iiniversally  adopted. 

In  his  first  classification,  published  in  1872,  Cohn  considered  the 
Bacteria  as  a  distinct  group  belonging  to  the  Algse,  and  divisible 
into  four  tribes,  including  six  genera  : — 

I.  Sphaerobacteria  .  .  globules  (Micrococcus). 

II.  Microbacteria  .  .  short  rods  (Bacterium). 

III.  Desmobacteria  .  .  long  rods  (Bacillus  and  Vibrio). 

IV.  Spirobacteria  .  .  spirals  (Sphirochseta  and  Spirillum). 

Cohn  noted,  in  spite  of  placing  them  with  the  Algae,  that  the 
absence  of  chlorophyll  connected  the  Bacteria  to  Fungi,  and  we  find 
Naegeli  subsequently  adopting  this  view,  and  employing  the  term 
Schizomycetes  or  Fission-fungi. 

Billroth,  in  1874,  disputed  the  division  into  species,  and  con- 
sidered that  all  the  forms  described  by  Cohn  were  but  developmental 
forms  of  one  micro-organism,  Coccobacteria  septica.  In  the  following 
year  Cohn  answered  the  criticism  of  Billroth,  and  produced  a  second 
classification,  in  which  he  still  maintained  that  distinct  genera  and 
species  existed.  Cohn  considered  the  genera  to  be  distinguished  by 
definite  differences  in  shape,  which  were  adhered  to  throughout  life, 
while  some  special  feature,  as  a  difference  in  size  or  physiological 
action,  or  some  minute  difference  in  form,  determined  the  various 
.species. 

The  second  classification  of  Cohn  (1875)  only  differed  from  the 
first  in  that,  instead  of  keeping  the  bacteria  as  a  separate  group,  he 
placed  them,  from  their  close  relationship  with  the  Phycochromacse, 
under  a  new  group,  the  Schizophytes,  and  added  the  genera  Lepto- 
thrix, Beggiatoa,  Crenothrix,  Sarcina,  Ascococcus,  Streptococcus, 
Myconostoc,  and  Streptothrix. 

Flugge  retained  the  term  Schizomycetes,  and  divided  them  thus  : — 


CLASSIFICATION   AND    DESCRIPTION   OF   SPECIES. 


477 


SCHIZOMYCETES   (FLtJGGE's    OKIGINAL    CLASSIFICATION). 

Isolated,  or  in  chains, 
or  united  in  amor- 
phous gelatinous 
families 


Micrococcus. 


Forming  gelatinous 
families  of  definite 
form. 


Colonies  solid,  filled 
with  cells 


Colonies,  with  simple 
layer  of  cells  at  the 
periphery  . 


In  large  numbers, 
in  irregular 
colonies  .  Ascococcus. 


In  small  but  defi- 
nite numbers,  in 
regular  groups  .  Sarcina. 


Clathrocystis* 


Short. 


Isolated,  or  in  small 
heaps  loosely  united, 
or  in  irregular  gela- 
tinous families 


Short, 
distinctly 
jointed 


Long. 


Threads 
isolated, 
i  n  t  e  r  -<; 
laced,  or 
in    bun- 
dles. 


Without 
ramifica- 
tions. 


Straight 
filaments. 


Long,      ^ 
notdis-JThin 
tinctly  j  Thick 
jointed.' 


Wavy,      or 


( Short  rigid 


Pseudo-ramifications 


.  Bacterium. 


.  Bacillus. 


Leptotlvrix . 


.  Threads  in  roundish  gelatinous  masses 


Spirochffte 
(vibrio}. 
Spirillum. 


fStreptotltnj'. 
\Clathrotlirli-. 


.  Myconostoc. 


478  SYSTEMATIC. 

The  belief  is  nevertheless  rapidly  gaming  ground  that  the  lowest 
forms  of  vegetable  life  cannot  be  divided  by  a  hard-and-fast  line 
into  a  series  with  chlorophyll  (Algse),  and  a  series  without  it  (Fungi), 
and  the  tendency  now  is  to  solve  the  difference  of  opinion  between 
Cohn  and  Nageli  by  following  the  example  of  Sachs,  and  amalga- 
mating the  two  series  into  one  group,  the  Thallophytes. 

Researches  by  competent  observers  have  more  recently  clearly 
demonstrated  that  several  micro-organisms  in  their  life  cycle  exhibit 
successively  the  shapes  characteristic  of  the  orders  of  Cohn. 

This  doctrine  of  pleomorphism,  now  widely  accepted,  was  dis- 
tinctly foreshadowed  in  a  publication  by  Lister  in  1873,  though 
this  memoir  contained  certain  conclusions  which  have  since  been 
abandoned.  Lister  described  forms  of  cocci,  bacteria,  bacilli,  and 
streptothrix  in  milk,  which  he  regarded  as  phases  of  the  same 
micro-organism,  Bacterium  lactis.  As  a  result  of  his  observations, 
Lister  remarks  that  "  any  classification  of  bacteria  hitherto  made 
from  that  of  Ehrenberg  to  that  of  Cohn  based  upon  absolute  mor- 
phological characters  is  entirely  untrustworthy."  To  Lankester, 
however,  belongs  the  credit  of  having  definitely  and  precisely  formu- 
lated this  doctrine.  In  a  paper,  also  published  in  1873,  Lankester 
observed  that  the  series  of  form-phases  which  he  had  discovered  in 
the  case  of  a  peach-coloured  bacterium  led  him  to  suppose  that 
the  natural  species  of  these  plants  were  "  within  the  proper  limits 
protean,  and  that  the  existence  of  true  species  of  bacteria  must  be 
characterised,  not  by  the  simple  form-features  used  by  Cohn,  but  by 
the  ensemble  of  their  morphological  and  physiological  properties  as 
exhibited  in  their  complete  life- histories."  Lankester  inferred  that 
these  phase-forms  were  genetically  connected,  in  that  they  all  pos- 
sessed the  common  characteristic  of  a  special  pigment,  bacterio- 
purpurin.  These  conclusions  were  vigorously  opposed  by  Cohn,  and 
doubt  still  remains  in  the  minds  of  some  as  to  whether  the  different 
forms  are  really  only  stages  in  the  life-history  of  a  single  species. 
Nevertheless  the  theory  of  pleomorphism  has  steadily  gained  ground 
ever  since. 

Cienkowski  and  Neelsen  worked  out  the  different  forms  assumed 
by  the  bacillus  of  blue  milk ;  Zopf  has  in  a  similar  manner  investi- 
gated Cladothrix,  Beggiatoa,  and  Crenothrix,  and  traced  out  various 
forms  (Fig.  196) ;  Yan  Tieghem  has  investigated  Bacillus  amylobacter 
with  a  similar  result;  Hauser  has  described  bacillar,  spirillar, 
spirulinar,  and  various  other  forms  in  the  Proteus  mirabilis  and 
Proteus  vulgaris.  These  facts  obviously  shake  the  very  foundation 
of  Cohn's  classification,  and  we  are  left  without  possessing  a  sound 


CLASSIFICATION   AND   DESCRIPTION   OF   SPECIES. 


479 


basis  for  classification  into  genera  or  species.  The  mode  of  repro- 
duction is  not  sufficiently  known  to  afford  a  better  means  for 
distinction  than  the  other  morphological  appearances  taken  alone ; 
nor  can  we  depend  upon  physiological  action,  which  is  held  by 
many  to  vary  with  the  change  of  form,  according  to  altered 
surroundings. 


FIG.  196. 

CLADOTHRIX  DICHOTOMA.— A.  Branched  Schizomycete  :  (a)  Vibrio-form  ;  (b)  Spiril- 
lum-form [slightly  magnified].  B.  Screw-form  at  the  ends :  (a)  Spirillum- 
form  ;  (6)  Vibrio-form.  C.  Very  long  Spirochseta-form.  D.  Branch  fragment, 
at  one  end  Spirillum-form,  at  the  other  Vibrio-form.  E.  Screw-form : 
(a)  Continuous ;  (6)  Composed  of  rods ;  and  (c)  Cocci.  F.  Spirochseta-form  : 
(a)  Continuous ;  (6)  Composed  of  long  rods ;  (c)  Short  rods ;  and  (d)  Cocci 
(Zopf). 

Zopf,  who  has  warmly  supported  the  pleomorphism  of  bacteria, 
has  suggested  as  a  result  of  his  investigations  a  division  of  the 
Schizomycetes,  Spaltpilze,  or  Fission -fungi,  into  the  following  four 
groups : — 


480  SYSTEMATIC. 


ZOPFS    CLASSIFICATION. 

GROUP  I.  COCCACEJE. — Possessing  (so  far  as  our  knowledge  at  present 

reaches)  only  cocci,  and  thread-forms  resulting  from  the  juxtaposition  of 

cocci.     The  fission  occurs  in  one  or  more  directions. 

Genus  I.  Streptococcus  (Chain-cocci). — Division  in  one  or  more  direc- 
tions. Individual  cocci  remain  united  together  to  form  chains. 

Genus  II.  Merismopedia  (Plate-cocci). — Divisions  in  two  directions, 
forming  lamellae  or  plates. 

Genus  III.  Sarcina  (Packet-cocci). — Division  in  three  directions,  form- 
ing colonies  in  cubes  or  packets. 

Genus  IV.  Micrococcus  (Mass-cocci). — Division  in  one  direction,  cocci 
after  division  remain  aggregated  in  irregular  clusters,  or  singly,  or 
in  pairs  or  in  chains  of  three  or  four  elements. 

Genus  Y.  Ascococcus  (Pellicle-cocci). — Like  micrococcus,  but  the  cocci 
grow  in  characteristic  gelatinous  pellicles. 

GROUP  II.    BACTERIACE^E. — Possessing  mostly  cocci,  rods  (straight 
or  bent),  and  thread-forms  (straight  or  spiral).     The  first  may  be  absent, 
and  the  last  possess  no  distinction  between  base  and  apex.     Division  (as 
far  as  is  known)  occurs  only  in  one  direction. 
Genus  I.     Bacterium. — Cocci  and  rods,  or  only  rods,  which  are  joined 

together  to  form  threads.     Spore-formation  absent  or  unknown. 
Genus  II.     Spirillum. — Threads  screw-form,  made  up  of  rods  (long  or 

short)   only,    or    of   rods    and    cocci.      Spore-formation   absent    or 

unknown. 
Genus  III.     Leuconostoc. — Cocci  and  rods.     Spore-formation  present  in 

cocci. 
Genus  IV.    Bacillus. — Cocci  and  rods,  or  rods  only,  forming  straight  or 

twisted  threads.     Spore-formation  present  either  in  rods  or  cocci. 
Genus  V.     Vibrio. — Threads  screw-form  in  long  or  short  links.     Spore- 
formation  present. 
Genus  VI.     Clostridium. — Same  as  bacillus,  but   spore-formation  takes 

place  in  characteristically  enlarged  rods. 

GROUP  III.     LEPTOTRICHEJE. — Possessing  cocci,  rods,  and  thread- 
forms   (which   show  a  distinction  between   base  and  apex).     The  last 
straight  or  spiral. 
Genus  I.     Crenothrix. — Threads  articulated  ;  cells  sulphurless  ;    habitat 

water. 
Genus    II.     Beggiatoa. — Threads    unarticulated ;    cells     with    sulphur 

granules  ;  habitat  water. 
Genus  III.     Phrngmidiothrix. — Threads  jointless  ;  successive  subdivision 

of  cells  is  continuous  ;  cells  sulphurless  ;  habitat  water. 
Genus  IV.     Leptothrix. — Threads  articulated  or  unarticulated  ;  successive 

subdivisions  of  cells  not  continuous  ;  cells  sulphurless. 

GROUP  IV.    CLADOTRICH^EE. — Possessing  cocci,   rods,   threads,  and 
spirals.     Thread-forms  provided  with  false  branchings. 
Genus :— Cladothrix. 


CLASSIFICATION   AND    DESCRIPTION    OF   SPECIES.  481 

Zopf,  however,  does  not  assert  that  all  the  fission-fungi  exhibit 
this  pleornorphism,  nor  does  he  pretend  that  his  classification  will 
include  all  the  micro-organisms  described.  Cohn,  on  the  other 
hand,  was  ready  to  admit  that  all  the  forms  described  by  him  were 
not  truly  independent  species.  De  Bary,  Hueppe,  Baumgarten,  and 
Fliigge  have  expressed  other  \iews  with  regard  to  the  classification 
of  bacteria. 

De  Bary  divides  them  into  two  great  groups — bacteria  which 
form  endospores,  and  bacteria  which  form  arthrospores.  This 
affords  but  little  practical  assistance,  though  regarded  by 
botanists,  from  a  scientific  standpoint,  as  a  step  in  the  right 
direction. 

Hueppe,  acknowledging  that  the  fructification  must  eventually 
be  made  the  basis  for  classification,  suggests  an  arrangement  for 
provisional  use  in  which  this  view  is  introduced  (p.  482). 

It  has  already  been  mentioned  that  the  production  of  arthrospores 
is  only  established  in  a  very  few  species.  Therefore,  we  are 
hardly  justified  in  assuming  that  all  bacteria,  the  spore-formation 
of  which  is  quite  unknown,  are  to  be  included  with  those  in  which 
this  kind  of  fructification  has  been  observed,  and  consequently  to 
distinguish  genera  on  the  same  grounds  may  be  considered,  to  say 
the  least,  somewhat  premature.  In  Baumgarten's  classification  the 
genus  bacterium  is  dispensed  with,  and  the  genera  divided  into  two 
groups,  the  monomorphic  and  the  pleomorphic. 

GROUP  I. — MONOMORPHIC. 

Genera. — Coccus. 
Bacillus. 
Spirillum. 

GROUP  II.— PLEOMORPHIC. 

Genera. — Spirulina. 
Leptothrix. 
Cladothrix. 

Fliigge  also,  in  his  revised  classification,  includes  the  genus 
bacterium  in  the  genus  bacillus.  The  new  classification  differs 
also  from  the  original  one  in  the  grouping  together  of  the  different 
species  according  to  the  character  and  behaviour  of  the  colonies 
in  nutrient  gelatine.  The  abolition,  in  Fliigge's  and  Baumgarten's 
classification,  of  the  genus  bacterium  is  no  doubt  owing  to  confusion 
having  arisen  from  the  distinction,  between  a  bacterium  and  a 
bacillus,  being  made  to  depend  upon  length.  Observers-  differed 
as  to  whether  a  rod  of  a  certain  length  ought  to  be  considered  a 

31 


482 


SYSTEMATIC. 


CLASSIFICATION   AND   DESCRIPTION   OF   SPECIES. 


483 


bacterium  or  a  bacillus.  To  meet  this  difficulty  a  rough-and- 
ready  rule  was  suggested— viz.,  that  a  rod  less  than  twice  its 
breadth  in  length  should  be  considered  as  a  bacterium,  and  other- 
wise a  bacillus.  But  this  purely  arbitrary  division  was  inadequate, 
from  the  fact  that  a  rod  at  one  stage  of  its  growth  or  under  certain 
conditions  might,  as  far  as  length  went,  truly  be  a  bacterium,  and 
under  other  circumstances  be  of  such  a  length  as  to  entitle  its  being 
considered  a  bacillus  (Fig.  197).  We  should  avoid  such  confusion 
if  we  followed  Zopf,  and  acknowledged  as  a  difference  between  a 
bacterium  and  a  bacillus  the  presence  or  absence  of  that  form  of 
spore-formation  now  distinguished  as  endogenous  spore -formation. 
We  might  then  conveniently  retain  this  generic  term,  to  include 
that  group  of  rod-forms  in  which  this  spore-formation  is  as  yet 


FIG.  197. — FKIEDLANDER'S  PNEUMOCOCCUS,    x   1500  (Zopf). 


unknown ;  moreover,  we  should,  by  so  doing,  with  one  or  two 
exceptions,  collect  together  those  short  rod-forms  which  appear  to 
link  the  simple  cocci  to  the  spore-bearing  rods  or  bacilli. 

The  grouping  together  of  the  different  species  according  to  the 
character  of  the  colonies  in  nutrient  gelatine  is  also  of  questionable 
advisability.  These  characters  can  hardly  be  considered  to  be  of 
sufficient  importance,  or  indeed  in  many  cases  to  be  sufficiently 
constant,  to  serve  by  themselves  for  this  purpose.  In  many  cases 
a  slight  variation  in  the  composition  of  the  nutrient  medium  may 
considerably  affect  the  appearances  of  the  colonies.  At  the  same 
time,  the  appearances  are  very  characteristic  of  certain  species  of 
bacteria,  and  in  some  cases  the  characters  of  the  colonies,  together 
with  the  characters  of  the  growth  in  test-tubes,  assist  us  in 
distinguishing  species  which  are  morphologically  similar,  as  in  the 
case  of  the  comma-bacilli  of  Tinkler  and  of  Koch. 


484  SYSTEMATIC. 

The  classification  of  Zopf  will  lead  the  investigator  to  work  upon 
the  same  lines,  and  by  tracing  the  life-history  of  individual  forms 
in  pure-cultivations  either  to  extend  the  work  of  establishing 
protean  species  or  to  restrict  the  doctrine  of  pleomorphism  to  a  few 
forms.  For  though  the  author  prefers  the  classification  proposed  by 
Zopf,  he  is  not  prepared  to  accept  his  views  entirely— for  instance, 
to  regard  the  bacterium  of  rabbit  septicaemia  as  a  micrococcus. 

Any  arrangement  at  present  can  only  be  considered  provisional, 
and  therefore  the  most  practical  classification  must  be  considered 
the  best.  In  fact,  much  more  investigation  is  required  before  we 
can  arrive  at  a  permanent  and  thoroughly  scientific  classification 
of  the  known  bacteria.  Many  bacteria  have  been  described  by 
different  observers  as  different  species  which  are  really  identical. 
Many  micro-organisms  have  been  described  and  named  as  new 
species  with  very  insufficient  investigation.  The  determination  of 
species  rests  upon  the  accumulated  evidence  afforded  by  a  thorough 
knowledge  of  their  life-history.  The  morphological  appearances 
under  different  conditions  must  be  carefully  studied,  the  presence 
or  absence  of  movement  and  of  spore-formation,  and  when  present 
the  exact  character;  the  appearances  of  colonies  and  of  test-tube 
cultivations  in  different  media  and  under  different  circumstances ; 
the  liquefaction  and  other  changes  in  nutrient  media ;  the  nature 
of  the  chemical  products,  if  any,  and  the  effect  on  the  living  animal 
of  the  bacterium  itself  and  of  its  products,  in  varying  doses,  must 
all  be  taken  into  account.  We  must  also  ascertain  whether  the 
bacterium  is  an  aerobe  requiring  the  presence  of  oxygen,  or  an 
anaerobe  growing  only  in  the  absence  of  it,  or  .a  facultative  anaerobe 
growing  equally  well  with  or  without  it ;  and  lastly,  we  must  know 
whether  the  bacterium  is  a  parasite  requiring  a  living  host,  or  a 
saprophyte  existing  on  dead  animal  or  vegetable  matter,  or  a 
facultative  parasite  capable  both  of  growing  in  the  living  animal 
and  of  leading  a  saprophytic  existence.  Several  writers  have 
classified  the  bacteria  which  have  been  described  hitherto  by  taking 
some  of  these  characters  into  account,  and  so  preparing  a  list  which 
is  convenient  for  the  purpose  of  bacteriological  diagnosis.  A  system 
of  this  kind  is  of  value  in  leading  investigators  to  supply  information 
which  is  wanting  in  order  to  verify  and  amplify  the  information 
upon  which  the  classification  is  based,  and  to  identify  species  which 
have  been  described  under  different  names. 


CLASSIFICATION    OF    SPECIES 
FOE    BACTEBIOLOGICAL    DIAGNOSIS 

COCCI. 

(A)   GELATINE   NOT  LIQUEFIED, 
(a)  Chromogenic. 

COLOUB.  HABITAT. 

Micrococcus  violaceus         .        .        .       Violet        .        .  .  Water. 

Micrococcus  carneus  ....       Flesh-red  .        .  .  Water. 

Micrococcus  cerasinus  siccus       .         .       Cherry-red        .  .  Water. 

Micrococcus  cinnabareus     .         .         .       Cinnabar-red    .  .  Air ;  water. 

Micrococcus  aurantiacus      .         .         .       Orange      .         .  .  Water. 

Micrococcus  vecsicolor        .        .        .       Yellow      ...  .  Water. 

Micrococcus  luteus      ....       Yellow      .         .  .  Water. 

Micrococcus  citreus     ....       Yellow      .         .  .  Water. 

Micrococcus  ochroleucus     .         .         .       Yellow      .         .  .  Water. 

Micrococcus  cereus  flavus  .         .         .       Yellow      .         .  .  Water. 

Micrococcus  agilis  citreus  .        .        .       Yellow      .        .  .  Air. 

Micrococcus  flavus  tardigradus  .         .       Yellow      .         .  .  Air ;  water. 

Staphylococcus  viridis  flavescens        .       Greenish- yellow  .  Lymph. 


(b)  Non-Chromogenic. 

Micrococcus  candicans 
Micrococcus  candidus 
Micrococcus  concentricus  . 
Micrococcus  fervidosus 
Micrococcus  cereus  albus  . 
Micrococcus  aquatilis 
Micrococcus  aquatilis  invisibilis 
Micrococcus  cumulatus  tenuis  . 
Micrococcus  rosettaceus 
Micrococcus  viticulosus 
Micrococcus  plumosus 
Micrococcus  amylivorus    . 
Micrococcus  acidi  lactici    . 
Micrococcus  ureae 
Micrococcus  gingivae  pyogenes 
Micrococcus  salivarius  septicus 
Micrococcus  of  Manfredi  . 
Micrococcus  in  pleuropneumonia 
Micrococcus  in  trachoma  . 
Haematococcus  bovis .... 
Diplococcus  albicans  tardissimus 


485 


Air ;  water. 
Water. 
Water. 
Water. 
Pus;  water. 
Water. 
Water. 
Nasal  mucu>. 
Water. 
Air;  water. 
Water. 
'  Pear-blight. 
Milk. 

Air;  urine. 
Pus. 
Saliva. 
Sputum. 
Lymph. 
Conjunctiva. 
Urine. 
Vaginal  secretion 


486 


CLASSIFICATION   OF  SPECIES 


Diplococcus  coryzse 

Pseudo-diplococcus  pneumonias 

Micrococcus  tetragenus 

Micrococcus  tetragenus  mobilis  ventriculi 

Pediococcus  cerevisiae 

Pediococcus  acidi  lactici 

Streptococcus  pyogenes 

Streptococcus  brevis 

Streptococcus  septicus 

Streptococcus  vermiformis 

Streptococcus  of  mastitis  in  cows 

Streptococcus  in  strangles 

Streptococcus  acidi  lactici 

(B)   GELATINE   LIQUEFIED. 

(a)  Chromogenic. 

COLOUR. 

Micrococcus  agilis       ....  Pink . 

Micrococcus  roseus      ....  Pink . 

Staphylococcus  pyogenes  aureus         .  Orange 

Micrococcus  in  pemphigus          .         .  Orange 

Staphylococcus  salivarius  pyogenes   .  Orange 

Micrococcus  fuscus      ....  Brown 

Micrococcus  flavus  desidens        .         .  Yellowish-brown 

Micrococcus  cremoides        .         .         .  Yellowish-white 

Micrococcus  botryogenus    .         .         .  Yellowish. 

Micrococcus  Finlayensis     .         .         .  Pale-yellow 

Staphylococcus  pyogenes  citreus        .  Yellow 

Micrococcus  citreus  liquefaciens         .  Yellow 

Micrococcus  flavus  liquefaciens  .         .  Yellow 

Micrococcus  tetragenus  versatilis       .  Yellow 

Diplococcus  flavus  liquefaciens  tardus  Yellow 

Diplococcus  subflavus         .        .        .  Yellow 

Diplococcus  citreus  conglomeratus     .  Yellow 

Diplococcus  luteus       ....  Yellow 

Diplococcus  roseus      ....  Pink . 

Diplococcus  fluorescens  fcetidus          .  Green 

(b)  Non-Chromogenic. 

Micrococcus  albus  liquefaciens 

Micrococcus  aerogenes 

Micrococcus  radiatus 

Micrococcus  fcetidus  .         .         .         .         .         .         . 

Micrococcus  in  Biskra-button  or  Pendjeh  sore 
Micrococcus  in  influenza    .         .        .         .         . 

Micrococcus  Freudenreichi        ...... 

Micrococcus  in  yellow  fever       ...... 

Micrococcus  lactis  viscosus        ...... 

Micrococcus  acidi  lactici  liquefaciens        .... 

Micrococcus  ureae  liquefaciens 

Micrococcus  in  gangrenous  mastitis  in  sheep   . 

Staphylococcus  pyogenes  albus 

Staphylococcus  pyosepticus 

Diplococcus  albicans  amplus 


HABITAT. 

Nasal  mucus. 

Meningitis. 

Sputum. 

Stomach. 

Beer ;  air. 

Malt ;  hay-dust. 

Pus. 

Saliva. 

Soil. 

Water. 

Pus. 

Pus. 

Milk. 


Water. 

Sputum. 

Pus. 

Bullaa. 

Saliva. 

Water. 

Air ;  water. 

Water. 

Equine  tumours. 

Yellow  fever. 

Pus. 

Eczema. 

Air ;  water. 

Blood. 

Eczema. 

Vaginal  mucus. 

Pus ;  air. 

Water. 

Air. 

Posterior  nares. 


Nasal  mucus. 

Intestine. 

Air ;  water. 

Nasal  mucus. 

Pus. 

Blood. 

Milk. 

Blood. 

Cream. 

Cheesy  butter. 

Urine. 

Milk. 

Pus. 

Pus. 

Vaginal  secretion. 


FOR   BACTERIOLOGICAL   DIAGNOSIS.  487 

HABITAT. 

Pediococcus  albus Water. 

Streptococcus  liquefaciens Blood. 

Streptococcus  septicus  liquefaciens Blood. 

Streptococcus  albus Water. 

Streptococcus  of  Mannaberg Urine. 

Streptococcus  coli  gracilis          . Faeces. 

(c)    NO    GROWTH    IN    GELATINE. 

Micrococcus  pneumonia  crouposse Saliva. 

Micrococcus  endocarditidis  rugatus  .         .         .         .         .         .  Heart. 

Nitromonas  of  Winogradsky     .......  Soil. 

Micrococcus  in  pemphigus Bullae 

Micrococcus  in  influenza    .        .        ...        .        .        .  Sputum. 

Micrococcus  gonorrhoeas     ........  Pus. 

Diplococcus  intercellularis  meningitidis Meningitis. 

Micrococcus  tetragenus  subflavus Nasal  mucus. 

Streptococcus  giganteus  urethras       .        ;        .        .        .        .  Urethra. 

Streptococcus  of  Bonome Meningitis. 

(D)    GROWTH    IN    GELATINE    UNDETERMINED. 

Ascococcus  Bilrothii Meat  infusion. 

Leuconostoc  mesenteroides        .......       Beet    juice ;    mo- 
lasses. 

Streptococcus  of  progressive  tissue  necrosis  in  mice         .        .  Putrid  blood. 

Micrococcus  pyogenes  tenuis Pus. 

Micrococcus  of  pyaemia  in  rabbits Meat  infusion. 

Micrococcus  of  progressive  abscess  formation  in  mice      .        .  Putrid  blood. 

Micrococcus  of  Forbes Cabbage       cater- 
pillars. 

Micrococcus  in  syphilis Blood 

Streptococcus  Havaniensis Liver. 

Streptococcus  perniciosus  psittacorum Blood  of  parrot. 

Streptococcus  bombycis Diseased     silk- 


PACKET    COCCI. 

(A)  GELATINE   NOT   LIQUEFIED. 

(a)  Non-Chromogenic. 

Sarcina  pulmonum Phthisical  sputum. 

Sarcina  ventriculi Stomach. 

(B)  GELATINE   LIQUEFIED. 

(a)  Chromogenic. 

COLOUR. 

Sarcina  mobilis Red  ....  Ascitic  fluid. 

Sarcina  rosea Red  ....  Air. 

Sarcina  flavea      .  Yellow      .        .        .  Beer. 

Sarcina  lutea Yellow      .        .        .  Air. 

Sarcina  aurantiaca      ....  Orange-yellow .        .  Air ;  water. 


488  CLASSIFICATION   OF  SPECIES 

(b)  Non-Chromogenic. 


Sarcina  alba 
Sareina  Candida 


Air  ;  water. 
Air  of  breweries. 


RODS. 

(I.)  AEROBES    OR    FACULTATIVE    ANAEROBES 

(A)  GELATINE   NOT   LIQUEFIED. 
(a)  Chromogenic. 

(A)   SPORE-FORMATION    PRESENT. 
(jO)   Motile. 

COLOUR. 

Bacillus  cyanogenus 
Bacillus  erythrosporus 


Bacillus  brunneus 


Bacilhis  rubefaciens 
Bacillus  rubescens 
Bacillus  fuscus  limbatus 
Bacillus  beroliniensis  Indicus 
Bacillus  cyanogenus 
Bacillus  aurantiacus 
Bacillus  fluorescens  aureus 
Bacillus  fluorescens  longus  . 
Bacillus  fluorescens  tenuis  . 
Bacillus  constrictus 
Bacillus  subflavus 
Bacillus  aureus     . 
Bacillus  flavescens 
Bacillus  heminecrobiophilus 
Bacillus  canalis  parvus 
Bacillus  fluorescens  putidus 
Bacillus  dentalis  viridans 
Bacillus  virescens 


Bacillus  latericeus 
Bacillus  spiniferus 
Bacillus  in  purpur; 
Bacillus  uceus 
Bacillus  in  cholera  in  ducks 
Bacillus  flavo-coriaceus 
Bacillus  striatus  flavis . 
Bacillus  fuscus 


Greyish-blue    . 

Blue  milk. 

Greenish-yellow 

Water. 

iarrhcea  (Lesage)    Green 

Intestine. 

(V)  Non-Motile. 

Brown 

Water. 

(B)  SPORE-FORMATION   UNKNOWN. 

O)  Motile. 

Pale-pink 

Water. 

Pale-pink 

Sewers. 

us      ...       Brown 

Rotten  eggs. 

[ndicus      .         .       Indigo-blue 

Water. 

ordaniensis        .       Bluish 

Sewers. 

Orange 

Water. 

ireus          .         .       Orange 

Water. 

ngus  .         .         .       Greyish-yellow 

Water. 

nuis  .         .         .       Greenish-yellow 

Water. 

Pale-yellow 

Water. 

Pale-yellow 

Water. 

Golden-yellow  . 

Water  ;  eczema. 

.       Yellow     . 

Swamp-  water. 

philus         .         .       Yellowish 

Caseous  glands. 

3         ...       Yellowish 

Sewer-water. 

itidus         :         .       Greenish  . 

Water. 

ans     .         .         .       Opalescent-green 

Carious  dentine. 

Green 

Sputum. 

(J)  Non-Motile. 

Brick-red 

Water. 

Greyish-yellow 

Eczema. 

emorrhagica      .       Greyish  -yellow 

Blood. 

Orange-yellow  . 

Water. 

ducks         .         .       Yellowish 

Blood. 

is                 .         .       Sulphur-yellow 

Water. 

Sulphur-yellow 

Nasal  mucus. 

Deep-yellow     . 

Water. 

m-liquefaciens  .       Greenish  -yellow 

Water. 

FOR   BACTERIOLOGICAL   DIAGNOSIS. 


489 


(b)  Non-Chromogenic. 

(A)  SPORE-FORMATION  PRESENT. 

(a)  Motile. 

Bacillus  putrificus  coli       .         .        .        .•       . 
Bacillus  septicus  vesica      .         .        ... 
Bacillus  in  cancer 


HABITAT. 

Human  faeces. 

Cystitis. 
Stomach. 


Non-Motile. 


Bacillus  acidi  lactici  (Hueppe)  . 
Bacillus  coprogenes  foetidus 
Bacillus  subtilis  similans   . 
Bacillus  epidermidis  . 
Bacillus  of  Colomiatti 


Sour  milk. 
Swine  measles. 
Human  faeces. 
Skin. 
Conjunctiva. 


(B)  SPORE-FORMATION  UNKNOWN. 


(a)  Motile. 


Bacillus  oedematis  aerobicus 
Bacillus  of  Fulles  (I.) 
Bacillus  stolonatus     . 
Bacillus  venenosus  brevis  . 
Bacillus  venenosus 
Bacillus  gracilis  anaerobiescens 
Bacillus  invisibilis 
Bacillus  albus     .... 
Bacillus  venenosus  invisibilis     . 
Bacillus  aquatilis  sulcatus 
Bacillus  argenteo-phosphorescens 
Bacillus  gliscrogenus  . 
Bacillus  cystiformis    . 
Bacillus  of  Guillebeau 
Bacterium  Zopfii 
Bacillus  ventriculi 
Bacillus  coli  communis 
Bacillus  cavicida         .        . 
Bacillus  of  Utpadel    . 
Bacillus  aerogenes 
Helicobacterium  aerogenes 
Bacterium  aerogenes . 
Bacillus  meningitidis  purulentse 
Bacillus  pyogenes  fcetidus 
Proteus  Zenkeri 
Bacillus  enteritidis     . 
Bacillus  hyacinthi  septicus 
Proteus  lethalis  .... 
Bacillus  endocarditidis  griseus  . 
Bacillus  of  Roth  (I.)  . 
Bacillus  Schafferi 
Bacillus  of  swine-plague     . 
Bacillus  typhi  abdominalis 
Bacillus  cavicida  Havaniensis    . 
Bacillus  cuniculicida  Havaniensis 


Earth. 

Earth. 

Water. 

Water. 

Water. 

Water. 

Water. 

Water. 

Water 

Water. 

Sea-water. 

Urine. 

Urine. 

Milk. 

Intestine  ;  air. 

Stomach  of  dogs. 

Intestine. 

Intestine. 

Intestine. 

Intestine. 

Intestine. 

Intestine. 

Pus. 

Pus. 

Putrid  substances. 

Poisonous  meat. 

Rotten  hyacinths. 

Septicaemia. 

Endocarditis. 

Old  rags. 

Cheese;  potato. 

Lymphatic  glands. 

Spleen  and  glands. 

Intestine. 

Intestine 


490  CLASSIFICATION   OF   SPECIES 

(&)  Non-Motile. 

HABITAT. 

Bacillus  of  Okada Dust. 

Bacillus  pyogenes  soli Earth. 

Bacillus  of  Fulles  (II.)       ........  Soil. 

Bacillus  candicans     .   • Soil. 

Bacillus  septicus  agrigenus Manured  soil. 

Bacillus  scissus Soil. 

Bacillus  ubiquitus Air ;  water. 

Bacillus  multipediculus     . Air;  water. 

Bacillus  albus  anaerobiescens    .         .         .         .         .         .         .  Water. 

Bacillus  Zurnianus Water. 

Bacillus  canalis  capsulatus Sewer-water. 

Bacillus  of  Roth  (II.)        .        .        . Old  rags. 

Bacillus  tenuis  sputigenus Sputum. 

Bacillus  crassus  sputigenus Sputum. 

Bacillus  coprogenes  parvus Faeces. 

Bacillus  of  Fiocca Saliva. 

Bacillus  striatus  albus        . Nasal  mucus. 

Bacillus  capsulatus  mucosus Nasal  secretion. 

Bacillus  pseudo-diphtheriticus Healthy  throat. 

Bacterium  ureas Urine. 

Bacillus  nodosus  parvus Healthy  urethra. 

Bacillus  oxytocus  perniciosus Milk. 

Bacillus  lactis  pituitosi Milk. 

Bacillus  limbatus  acidi  lactici Milk. 

Bacillus  ovatus  minutissimus Eczema. 

Bacillus  of  Belfanti  and  Pascarola Pus. 

Bacillus  of  Tommasoli Hair  with  sycosis. 

Bacillus  capsulatus    .........  Blood. 

Bacillus  of  purpura  haemorrhagica  (Babes)       ....  Blood, 

Bacillus  of  purpura  haemorrhagica  (Kolb)         ....  Blood. 

Bacillus  septicaemiae  haemorrhagicae  ......  Blood. 

Proteus  capsulatus  septicus       .......  Blood. 

Bacillus  acidiformans Liver. 

Bacillus  coli  similis    .  Liver. 

Bacillus  hepaticus  fortuitus Liver. 

Bacillus  filiformis  Havaniensis Liver. 

Bacillus  of  Martinez  .........  Liver. 

Bacillus  diphtheriae  columbrarum Diphtheritic  de- 
posit. 

Bacillus  diphtheriae Diphtheritic 

throat. 

Bacillus  of  Schimmelbusch Cancrum  oris. 

Bacillus  capsulatus  Smithii Intestine. 

Bacillus  erysipelatis  suis Blood. 

Bacillus  in  rhinoscleroma Rhinoscleroma. 

Bacillus  of  Friedliinder Sputum. 

Bacillus  pneumosepticus Septic  pneumonia. 

Bacillus  endocarditidis  capsulatus Endocarditis. 

Bacillus  septicus  keratomalaciae Internal  organs. 

Bacillus  of  intestinal  diphtheria  in  rabbits       ....  Intestine. 

Bacillus  of  acne  contagiosa  of  horses Pus. 

Bacillus  pseudo-tuberculosis Internal  organs. 

Bacterium  tholcEideum Intestine. 

Bacillus  gallinarum Blood. 


FOR   BACTERIOLOGICAL    DIAGNOSIS. 


491 


Bacillus  argenteo-phosphorescens     . 
Bacillus  smaragdino-phosphorescens 
Bacillus  phosphorescens  gelidus 
Proteus  hominis  capsulatus 
Bacillus  of  grouse  disease. 
Bacillus  lactis  aerogenes    . 


HABITAT. 

Fish. 

Fish. 

Cuttlefish. 

Blood. 

Blood. 

Intestine. 


(A)    GELATINE    LIQUEFIED, 
(a)  Chromogenic. 

(A)    SPOKE-FORMATION    PRESENT. 


O) 

Motile. 

COLOUR. 

Bacillus  violaceus 

Deep-violet       .        .       Water. 

Bacillus  in  disease  of  bees  (Canestrini) 

Pink  ....       Larvae  of  bees. 

Bacillus  in  "  red-cod  " 

Red  .        .        .        .      Salted  codfish. 

Bacillus  inesentericus  ruber        .        . 

Reddish  -yellow        .      Potatoes. 

Bacillus       fluorescens      liquefaciens 

minutissimus    .        .        ... 

Greenish-yellow        .       Skin. 

(B)    SPORE-FORMATION    UNKNOWN. 

(a) 

Motile. 

Bacillus  ianthinus        .         .        . 

Bluish-violet     .        .       Water. 

Bacillus  violaceus  Laurentius     . 

Deep-violet       .        .       Water. 

Bacillus  lividus    .         .         .         .        .  .  - 

Violet-black      .        .       Water. 

Bacillus  carnicolor      .... 

Dark  flesh  -colour      .       Water. 

Bacillus  rubidus  

Brownish-red    .        .       Water. 

Bacillus  Indicus  

Sealingwax-red         .       Intestine. 

Bacillus  rosaceus  metalloides 

Magenta-red     .        .       Water. 

Bacillus  ochraceus       .... 

Yellow      .        .        .       Water. 

Bacillus  citreus  cadaveris   . 

Yellow      .        .        .       Blood. 

Bacillus  buccalis  minutus  . 

Yellow      .        .        .       Saliva. 

Bacillus  arborescens    .... 

Yellow      .        .        .       Water. 

Bacillus  fulvus     ..... 

Yellow      .        .        .       Water. 

Bacillus  plicatilis         .... 

Yellowish          .        .       Water. 

Bacillus  pyocyaneus   .... 

Yellowish-green        .       Pus. 

Bacillus  fluorescens  liquefaciens 

Greenish-yellow\       .      Water. 

Bacillus  cyanofuscus  .... 

Greenish  -brown        .       Cheese  ;  glue. 

Bacillus  fluorescens  nivalis 

Bluish  -green     .        .       Water. 

Bacillus  chromo-aromaticus 

Green  or  brown        .       Intestine. 

Bacillus  viscosus.         .... 

Green        .        .         .      Water. 

Bacillus  pyocyaneus    .... 

Green        .        .        .       Pus. 

(&)  Non-Motile. 

Bacillus  coeruleus         .... 

Blue  ....      Water. 

Bacillus  glaucus  

Grey.        .        .        .      Water. 

Bacillus  membranaceus  amethystinus 

Violet        .        .        .       Water. 

Bacillus  lactis  erythrogenes 

Red  .        .        .        .      Milk. 

Bacillus  mycoides  roseus     . 

Red  ....      Soil. 

492 


CLASSIFICATION   OF   SPECIES 


COLOUR. 

Bacillus  prodigiosus    ....  Blood-red  . 

Bacillus  hydrophilus  fuscus         .        .  Yellow 

Bacillus  cuticularis     ....  Yellow 

Bacillus  helvolus          ....  Yellow 

Bacillus  tremelloides  ....  Yellow 

Ascobacillus  citreus    ....  Yellow 
Bacillus         argenteo-phosphorescens 

liquefaciens Yellowish 

Bacterium  termo  (Vignal)  .         .         .  Yellowish 

Bacillus  smaragdinus  fcetidus     .        .  Green 


Air. 

Frog's  lymph. 

Water. 

Water. 

Water. 

Skin. 

Sea-water. 

Saliva. 

Ozaena. 


(b)  Non-Chromogenic. 

(A)    SPORE-FORMATION    PRESENT. 


Motile. 


Bacillus  inflatus          .        .        . 
Urobacillus  Freudenreichi 
Bacillus  mycoides 
Bacillus  ramosus         .        . 
Bacillus  gracilis .         .        .,        , 
Bacillus  circulans 
Urobacillus  Duclauxi 
Urobacillus  Maddoxi 
Bacillus  limosus .... 
Bacillus  butyricus 
Bacillus  Hessii    .... 
Bacillus  lactis  albus   . 
Bacillus  liodermos 
Urobacillus  Pasteuri  . 
Bacillus  meaentericus  fuscus 

Bacillus  mesentericus  vulgatus  . 
Bacillus  of  potato  rot 
Bacillus  maidis  .... 
Bacillus  megatherium 
Bacillus  tumescens      . 
Bacillus  subtilis . 
Bacillus  subtilis  similis 
Bacillus  vacuolosis     . 
Bacillus  of  Scheurlen 
Bacillus  alvei 


Air. 

Air ;  dust ;  sewers. 

Soil;  water. 

Soil ;  water. 

Water. 

Water. 

Water. 

Water. 

Sea-dredgings. 

Milk. 

Milk. 

Milk. 

Milk. 

Urine. 

Potato;  dust; 

water. 

Potato;  water,  etc. 
Rotting  potatoes. 
Maize  infusion. 
Boiled  cabbage. 
Beet-root. 
Dust ;  water ;  soil. 
Liver. 
Liver. 

Cancerous  tissues. 
Larvae  of  bees. 


(b)  Non-Motile. 


Bacillus  aerophilus     . 
Bacillus  implexus 
Bacillus  filiformis 
Bacillus  vermicularis 
Bacillus  incanus 
Bacillus  inunctus 
Bacillus  granulosus    . 
Bacillus  carotarum     , 


Air. 
Water. 

Water. 
Water. 

Swamp-water. 
Swamp- water. 
Sea-dredgings. 
Boiled  carrot. 


FOR   BACTERIOLOGICAL   DIAGNOSIS.  493 


Bacillus  brassicae Cabbage  infusion. 

Bacillus  in  gangrene Senile  gangrene. 

Bacillus  of  Letzerich  .........  Urine. 

Bacillus  anthracis Blood. 


(B)    SPORE-FORMATION    UNKNOWN, 
(a)  Motile. 

Bacillus  pestifer         .        . Air. 

Bacillus  diffusus         .  .......  Soil. 

Bacillus  gasoformans          .        .        ...        .        .         .  Water. 

Bacillus  liquidus Water. 

Bacillus  guttatus        .        .        .        .     '. Water. 

Bacillus  liquefaciens  .        .        .        ".  ; Water. 

Bacillus  radiatus  aquatilis         .         <        .        .         .         .         .  Water. 

Bacillus  nubilis Water. 

Bacillus  albus  putidus        .        .  .        .        .        .        .  Water. 

Bacillus  hyalinus        .         . Water. 

Bacillus  vermiculosus         ....        .        .        .        .  Water. 

Bacillus  dehcatulus     .         .         .         .        .         .         .         .         .  Water. 

Bacillus  punctatus Water. 

Bacillus  reticulans Water. 

Bacillus  figurans  (Vaughan)       . Water. 

Urobacillus  Schutzenbergi Water. 

Bacillus  devorans Water. 

Bacillus  venenosus  liquefaciens Water. 

Bacillus  aquatilis Water. 

Proteus  sulphureus Water. 

Bacillus  stoloniferus Swamp-water. 

Bacillus  phosphorescens  Indicus Sea-water. 

Bacillus  phosphorescens  indigenus Sea-water. 

Bacillus  cyaneo-phosphorescens Sea-water. 

Bacillus  litoralis Sea-dredgings. 

Bacillus  halophilus Sea-dredgings. 

Bacillus  superficialis Sewage. 

Bacillus  cloacae  .  Sewage. 

Proteus  microsepticus Uterine     dis- 
charges. 

Proteus  vulgaris ..........  Putrid  substances. 

Proteus  mirabilis Putrid  substances. 

Proteus  septicus .  Septicaemia. 

Bacillus  foetidus  ozsenae Nasal  mucus. 

Bacillus  septicus  ulceris  gangraenosi Blood  and  organs. 

Bacillus  albus  cadaveris Blood. 

Bacillus  of  Guillebeau Milk. 

Bacillus  Havaniensis  liquefaciens Skin. 

Bacillus  carabiformis Stomach. 

Bacillus  of  Schou Rabbit. 

Bacillus  leporis  lethalis Liver. 

Bacillus  liquefaciens  communis Liver. 

(*)  Non-Motile. 

Bacillus  buccalis  fortuitus Saliva. 


494  CLASSIFICATION   OF   SPECIES 


HABITAT. 

Bacillus  ulna  (Vignal) Saliva.  p 

Leptothrix  buccalis  (Vignal) Mouth. 

Bacillus  varicosus  conjunctive Conjunctiva. 

Bacillus  gingivae  pyogenes Alveolar  abscess. 

Bacillus  pulpse  pyogenes .         .  Gangrenous  tooth- 
pulp. 

Bacillus  graveolens Skin  of  feet. 

Pneumo-bacillus  liquefaciens  bo  vis Lung. 


(c)   NO  GROWTH  IN  GELATINE. 

(A)  SPORE-FORMATION   PRESENT. 

O)  Motile. 

Bacillus  ulna White  of  egg. 

Bacillus  in  putrid  bronchitis Sputum. 

Bacillus  mallei Glandered  tissue. 

(ft)  Non-Motile. 

Bacillus  in  erythema  nodosum Blood. 

Bacillus  tuberculosis Tubercular  tissue, 

(B)  SPORE-FORMATION  ABSENT. 

Bacillus  sanguinis  typhi Blood. 

Bacillus  septicus  acuminatus Septic  infection, 

Bacillus  necrophorus Condyloma. 

(c)  SPORE-FORMATION  NOT   STATED. 

Bacillus  allantoides ^jr 

Bacillus  nitrificans gojj 

Bacillus  in  measles Blood 

Bacillus  in  ophthalmia       ....                 .'.'.'  Conjunctiva. 

(D)  GROWTH  IN  GELATINE   NOT   STATED. 

Bacillus  senilis Blood. 

Bacillus  leptosporus Air. 

Bacillus  allii       .         . .'  Putrefying  onions. 

Bacillus  mdigogenus Infusion  of  indigo. 


II.     ANAEROBES. 
(A)  GELATINE    LIQUEFIED. 

(A)    SPORE-FORMATION    PRESENT. 
(«)   Motile. 

Bacillus  rubellus Dust 

Bacillus  butyricus  (Botkin)        ....'.'.'        .'       Milkjwater;    dust 
Clostridmm  fcetidum Earth 


FOR   BACTERIOLOGICAL   DIAGNOSIS.  495 

HABITAT. 

Bacillus  radiatus Earth. 

Bacillus  liquefaciens  magnus Earth. 

Bacillus  spinosus Earth. 

Bacillus  thalassophilus Sea-dredgings. 

Bacillus  of  symptomatic  anthrax Tissues  (quarter-ill). 

Bacillus  cedematis  maligni Lymph. 

Bacillus  tetani Wounds ;  earth. 

(&)  Non-Motile. 

Bacillus  liquefaciens  parvus       .......  Earth. 

Bacillus  anaerobicus  liquefaciens Yellow  fever. 


(B)   GELATINE  NOT  LIQUEFIED. 

(A)  SPORE-FORMATION  PRESENT. 

O)  Motile. 

Bacillus  amylozyma Water. 

Bacillus  solidus .         .       Earth. 

Bacillus  polypiformis Earth. 

(&)  Non-Motile. 
Bacillus  muscoides Earth. 

(B)  SPORE-FORMATION  ABSENT. 

(&)  Non-Motile. 
Bacillus  aerogenes  capsulatus Blood. 

(c)  GROWTH  IN  GELATINE  NOT  STATED. 

(A)  SPORE-FORMATION   PRESENT. 
(a)  Motile. 

Bacillus  butyricus Vegetable      infu- 
sions, etc. 

(B)  SPORE-FORMATION  UNKNOWN. 
Bacillus  cadaveris       .        *• Liver. 

CURVED    RODS. 

(A)   GELATINE  NOT   LIQUEFIED. 

(a)  Chromogenic. 

O)  Motile. 

COLOUR. 

Spirillum  rubrum        ....     Deep-red     .        .        .      Putrid  mouse. 

(b)  Non-Chromogenic. 

Spirillum  suis Intestine  of  swine. 


496  CLASSIFICATION    OF   SPECIES 


Spirillum  concentricum      ...  ....       Putrefying  blood. 

Spirillum  saprophiles .        .       Hay -infusion; 

sewage. 

(i)  Non- Motile. 
(a)  Chromogenic. 


COLOUR. 


Spirillum  flavescens     ....     Yellowish-green          .  Sewers. 

Spirillum  flavum          ....     Ochre-yellow      .         .  Sewers. 

Spirillum  aureutn         ....     Orange-yellow    .         .  Sewers. 

(b)  Non-Chromogenic. 

Spirillum  linguae Deposit  on  tongue. 

Spirillum  nasale Nasal  mucus. 

GELATINE    LIQUEFIED, 
(a)  Non-Chromogenic. 

(a)  Motile. 

Spirillum  cholerse  Asiaticae        .         .   " Intestine. 

Spirillum  of  Finkler  and  Prior Intestine. 

Spirillum  Metchnikovi       .         .         ...         .         .         .  Intestine  of  fowls. 

Spirillum  of  Miller Carious  teebh. 

Spirillum  of  Sanarelli Water. 

Spirillum  tyrogenum Old  cheese. 

Spirillum  marinum    .         .         . Sea-dredgings. 


NO    GROWTH    IN    GELATINE,    OR    UNDETERMINED. 

(a)  Motile. 

Spirillum  Obermeieri          .         . Blood. 

Spirillum  anserum Blood  of  geese. 

Spirillum  undula Putrid  infusions. 

Spirillum  sputigenum .         .  Gums. 

Spirillum  serpens        .........  Stagnant  water. 

Spirillum  tenue Putrid  infusions. 

Spirillum  volutans Swamp-water. 

Spirillum  plicatile Swamp-water. 

(&)  Non-Motile. 

Spirillum  dentium Gums. 

Spirillum  sanguineum        . Brackish  water. 

BRANCHING    FILAMENTS. 

Streptothrix  actinomycotica. 
Streptothrix  alba. 
Streptothrix  liquefaciens. 
Streptothrix  musculorum  suis. 
Streptothrix  Hofmanni. 
Streptothrix  farcinica. 
Streptothrix  asteroides. 
Streptothrix  carnea. 


FOR   BACTERIOLOGICAL    DIAGNOSIS.  497 

Streptothrix  aurantiaca. 
Streptothrix  chromogenes. 
Streptothrix  odorifera. 
Streptothrix  violacea. 
Streptothrix  Forsteri. 
Streptothrix  madurae. 


NOT    CLASSIFIED. 

Bacillus  indigonaceus. 

Bacillus  proteus  fluorescens. 

Beggiatoa  alba. 

Beggiatoa  mirabilis. 

Beggiatoa  roseo-persicina. 

Cladothrix  dichotoma. 

Cladothrix  invulnerabilis. 

Crenothrix  Kuhniana. 

Diplococcus  citreus  liquefaciens. 

Leptothrix  buccalis. 

Leptothrix  gigantea. 

Micrococcus  aquatilis  invisibilis. 

Micrococcus  crepusculum. 

Micrococcus  foetidus. 

Micrococcus  Havaniensis. 

Micrococcus  of  septicaemia  in  rabbits. 

Monas  Okenii. 

Monas  vinosa. 

Monas  Warmingii. 

Myconostoc  gregarium. 

Rhabdomonas  rosea. 

Sarcina  hyalina. 

Sarcina  intestinalis. 

Sarcina  litoralis. 

Sarcina  Reitenbachii. 

Sarcina  urinae. 

Sphaerotilus  natans. 

Spirillum  attenuatum. 

Spirillum  leucomelaneum. 

Spirillum  rosaceum. 

•Spirillum  Rosenbergii. 

Spirillum  rufum. 

Spiromonas  Cohnii. 

Spimmonas  volubilis. 

Streptococcus  cadaveris. 

Streptococcus  flavus  desidens. 

Vibrio  rugula. 


DESCRIPTION    OF    SPECIES    AEEANGED    FOE 
REFEBENCE   IN  ALPHABETICAL   OBDEB. 


Ascococcus  Billrothii.— Small 
globular  cocci,  united  into  charac- 
teristic colonies. 

They  form  on  the  surface  of 
nourishing  fluids  a  cream-like  skin, 
divisible  into  an  enormous  number 
of  globular  or  oval  families.  Each 
family  is  surrounded  by  a  thick 
capsule  of  cartilaginous  consistency. 
In  a  solution  containing  acid  tar- 
trate  of  ammonia  the  fungi  generate 
butyric  acid,  and  change  the  origin- 
ally acid  fluid  into  an  alkaline  one. 


FIG.  198. — Ascococcus  BILLROTHII 
(Cohn). 

They  were  first  observed  on  putrid 
broth,  and  later  on  ordinary  nourish- 
ing solutions ;  they  also  readily  de- 
velop upon  damp  slices  of  boiled 
roots,  carrots,  beetroots,  etc. 

Ascobacillus  citreus  (Unna, 
Tommasoli). —  Rods  sometimes 
curved,  T3  p.  in  length,  '3  p  in  width, 
singly,  in  pairs,  and  masses.  The 


colonies  develop  slowly,  and  are 
yellowish  in  colour. 

The  cocci  inoculated  in  the  depth 
of  gelatine  form  small  colonies  in 
the  track  of  the  needle,  and  a  slimv 
pale-yellow  growth  on  the  surface"; 
liquefaction  sets  in  slowly. 

On  agar  the  growth  is  gelatinous, 
and  orange  in  colour,  and  rapidly 
extends  over  the  surface. 

On  potato  the  growth  is  abundant, 
and  pale  yellow. 

They  were  isolated  from  the  skin 
in  eczema  seborrhoeicum. 

Bacillus  acidiformans  (Stern- 
berg). — Short  rods,  1  -5  to  3  p  in 
length,  1'2  /i  in  width,  and  filaments 
5  to  10  p. 

Colonies  circular  ;  iridescent  by 
reflected  light. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  freely  in  the  track  of 
the  needle,  and  form  a  hemispherical 
mass  on  the  surface.  They  produce 
gas  bubbles. 

On  agar  the  growth  is  milk-white, 
and  the  jelly  becomes  strongly  acid. 
On  potato  the  growth  is  abundant. 

In  broth  with  5  per  cent,  glycerine 
they  produce  opacity  and  a  copious 
viscid  deposit,  and  the  surface  is 
covered  with  gas  bubbles. 

Injected  into  the  peritoneal 
cavity  of  rabbits  and  guinea-pigs, 
they  produce  death  in  twenty-four 
hours. 

They  were  isolated  from  the  liver 
in  a  fatal  case  of  yellow  fever. 

Bacillus  acidi  lactici  (Hueppe). 
—Rods  1  to  2'8  /LI  long,  and  '3  to 
'4  /x  wide,  and  thread  forms.  Spore- 
formation  present.  In  gelatine 


498 


DESCRIPTION   OF   SPECIES. 


cultures  the  breadth  of  the  rods  is 
diminished.  They  grow  best  be- 
tween 35°  and  42°  C.,  and  cease 
under  10°  C.  Over  45'5°  C.  they 
no  longer  produce  acidity. 

Whitish  colonies  appear  on  the 
second  day. 

In  gelatine  a  delicate  growth 
appears  along  the  whole  track  of 
the  needle,  with  spherical  forms 
here  and  there. 

In  milk  they  produce  lactic  acid 
and  the  casein  is  precipitated. 

Bacillus  aerogenes  (Miller): — 
Small  rods  varying  in  length. 
Colonies  white  or  yellowish- white  ; 
concentric. 

In  the  depth  of  gelatine  they 
produce  a  yellowish  filament,  and 
on  the  surface  a  grey  patch  with 
dentated  periphery  ;  later  the  fila- 
ment is  brown. 

On  potato  the  growth  is  yellowish 
and  dry. 

They  were  isolated  from  the  in- 
testine in  health. 

Bacillus  aerogenes  capsulatus 
(Welch). — Rods  straight  or  slightly 
curved,  3  to  6  p.  ;  threads  and 
chains  ;  capsulated. 

Colonies  on  agar  greyish- white, 
with  hairy  processes. 

They  peptonise  gelatine  and  pro- 
duce gas.  Broth  becomes  turbid, 
and  there  is  an  abundant  sediment. 
Milk  is  coagulated.  Cultures  have 
a  faint  smell  of  glue. 

Injected  into  rabbits  they  pro- 
duce gas  in  the  blood  and  internal 
organs. 

They  were  isolated  from  a  patient 
after  death,  with  blood-vessels  full 
of  gas. 

Bacillus  aerophilus  (Liborius). 
— Rods  and  filaments. 

Colonies  punctiform  ;  greyish- 
yellow. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  produce  a  funnel  of 
liquefied  jelly,  with  flocculi  in  the 
lower  part. 

On  potato  they  form  a  smooth 
yellowish  layer. 

They  were  isolated  from  con- 
taminated cultures. 

Bacillus  albus  (Eisenberg).— 
Rods  and  chains. 


Colonies  circular,  white. 

In  gelatine  the  bacilli  grow  in  the 
track  of  the  needle,  and  form  a 
white  hemispherical  mass  on  the 
free  surface. 

On  agar  the  growth  is  pure  white, 
and  on  potato  yellowish- white. 

They  occur  in  water. 

Bacillus  albus  anaerobiescens 
(Vaughan). — Short  rods. 

Colonies  circular,  yellowish- 
brown. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  in  the  track  of  the 
needle,  and  on  the  free  surface. 

On  agar  the  growth  is  pure  white, 
and  on  potato  yellowish-white. 

They  occur  in  water. 

Bacillus  albus  cadaveris 
(Straussmann  and  Strieker). — Rods 
'2-5  n  in  length,  '75  /z  in  width,  and 
filaments. 

Colonies  yellowish  ;  circular,  and 
later  radiated. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  a  funnel  of  lique- 
fied gelatine  with  a  thick  deposit. 

On  agar  there  is  an  abundant 
white  growth. 

On  potato  the  growth  is  white  or 
yellowish-white,  and  colours  the 
potato  in  the  vicinity  bluish-brown. 
The  cultures  have  a  putrefactive 
odour. 

Mice  inoculated  subcutaneously 
die  in  six  hours,  and  guinea-pigs  in 
twenty-four. 

They  were  isolated  from  putrid 
human  blood. 

Bacillus  albus  putidus  (De 
Bary). — Rods  and  filaments.  -:J 

Colonies  circular  and  brownish. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  rapid  lique- 
faction. 

On  agar  and  potato  the  growth 
is  slimy.  Cultures  develop  a  strong 
putrefactive  odour. 

They  occur  in  water. 

Bacillus  allantoides  (L.  Klein), 
— Rods  2  to  2*5  /u  in  length,  '5  p.  in 
width,  and  in  chains.  The  rods 
develop  cocci-forms  united  by  a 
gelatinous  substance  into  zooglcea 
masses.  They  were  isolated  from 
a  contaminated  culture. 

Bacillus  allii  (Griffiths).— Rods 


oOO 


DESCRIPTION   OF   SPECIES. 


5  to  7  p  in  length,  2 -5  /x  in  width, 
singly,  in  pairs,  and  zoogloea. 

On  agar  they  produce  a  bright 
green  film,  and  cultures  are  said  to 
emit  traces  of  sulphuretted  hydro- 
gen. 

They  were  isolated  from  putrid 
onions. 

Bacillus  alvei  (p.  470). 

Bacillus  amylozyma  (Perdrix). 
— Rods  2  to  3  /u,  in  length  and  "5  p 
in  width,  in.  pairs,  and  in  chains. 
They  are  anaerobic. 

Colonies  white,  and  producing  gas 
bubbles. 

On  potato  in  an  atmosphere  of 
hydrogen  the  bacilli  partly  liquefy 
it,  and  there  is  abundant  formation 
of  gas. 

They  ferment  sugar  and  starch. 

Bacillus  anaerobicus  lique- 
faciens  (Sternberg). — Slender  rods, 
about  '6  n  in  diam.,  in  pairs,  and 
in  filaments. 

Colonies  granular  and  white  ;  sur- 
rounded by  liquefied  gelatine. 

They  grow  along  the  track  of 
the  needle  when  inoculated  in  the 
depth  of  agar. 

They  were  isolated  from  the  in- 
testine in  a  fatal  case  of  yellow 
fever. 

Bacillus  anthracis  (p.  192). 

Bacillus  aquatilis  (Frankland). 
— Rods  2 -5  [JL  in  length,  and  filaments 
17  fj.  or  longer.  They  resemble 
Bacillus  arborescens. 

Colonies  after  liquefaction  of  the 
gelatine  have  a  yellowish-brown 
nucleus  from  which  proceed  twisted 
strands  of  filaments. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  in  the  track  of  the 
needle  is  at  first  almost  invisible, 
later  liquefaction  occurs. 

On  agar  the  growth  is  shining 
and  yellowish. 

Broth  becomes  turbid,  and  a 
sediment  forms  at  the  bottom  of 
the  tube. 

On    potato    there    is  a    slightly 
yellowish  streak. 
They  occur  in  water. 
Bacillus  aquatilis  fluorescens 
(Lustig). — Short    thin    rods    with 
rounded  ends.     Non-motile. 

Colonies  fern-like  and  iridescent. 


Compare  Eisenberg's  Bacillus fluo- 
rcscens  non-liqiiefaciens. 

Bacillus  aquatilis  graveolens 
(Tataroff). — Slender  rods  1-3  /A  in 
length.  They  rapidly  liquefy  gela- 
tine and  produce  an  odour  like  that 
of  perspiration  from  the  feet. 

They  occur  in  water. 

Bacillus  aquatilis  sulcatus 
(Weichselbaum),  No.  I. — Rods  mor- 
phologically, and  in  cultures  resem- 
bling Bacillus  typhosus. 

Colonies  in  gelatine  exhibit  lines 
and  furrows. 

The  growth  on  the  surface  of 
gelatine  is  said  to  be  greater  than  in 
cultures  of  Bacillus  typhosus  grown 
for  comparison. 

They  occur  in  water. 

No.  II. — Rods  also  resembling  in 
morphology  and  cultivation  the 
Bacillus  typhosus. 

Colonies  are  said  to  be  thicker  than 
those  of  No.  I.,  and  not  dentated. 

The  growth  on  potatoes  is  yel- 
lowish-brown, and  emits  a  faint 
odour  of  urine. 

They  occur  in  water. 

No.  III. — Very  short  rods. 

Colonies  show  lines  and  furrows, 
and  are  yellowish. 

On  the  surface  of  gelatine  the 
growth  develops  as  a  thin  whitish 
film. 

On  agar  the  growth  is  white  and 
abundant. 

On  potato  the  growth  is  yellow. 

They  were  isolated  from  water. 

No.  IV. — Rods  and  filaments. 

Colonies  circular  and  bluish. 

On  the  surface  of  gelatine  the 
growth  is  greyish-white,  and  on 
agar  there  is  a  similar  appearance. 

They  do  not  grow  on  potato. 

They  occur  in  water. 

No.  V. — Rods  rather  thicker  than 
those  of  Bacillus  typhosus. 

Colonies  similar  to  those  of  No.  I. 

The  growth  on  the  surface  of 
gelatine  is  yellow. 

On  agar  the  growth  is  viscid  and 
yellow,  and  on  potato  the  growth  is 
faintly  yellow  and  the  surrounding 
medium  stained  bluish-grey. 

They  occur  in  water. 

Bacillus  arborescens  (Frank- 
land). — Rods  2'5  p  in  length,  and 


DESCRIPTION   OF   SPECIES. 


501 


•5  p.  in  width,  singly,  in  pairs,  and 
in  short  chains,  and  filaments. 

Colonies  throw  out  delicate 
branches  with  a  highly  characteristic 
appearance  ;  the  gelatine  slowly 
liquefies,  the  nucleus  of  the  colony 
becomes  yellow,  and  the  periphery 
iridescent. 

.  Inoculated  in  the  depth  of  gela- 
tine the  bacilli  form  a  cloudiness  in 
the  track  of  the  needle,  and  an 
iridescent  layer  on  the  surface  with 
central  depression  of  the  gelatine 
and  commencing  liquefaction. 
Later  the  liquefaction  produces 
a  funnel,  and  there  is  a  yellow 
deposit. 

On  the  surface  of  agar  the  layer 
is  a  dirty  orange  colour. 

On  potato  the  growth  is  orange 
red  with  irregular  protuberances, 
and  limited  in  growth. 

They  occur  in  water. 

Bacillus  argenteo-phosphores- 
cens  (Katz),  No.  I.— Rods  slightly 
curved  with  pointed  ends,  2'5  ^  in 
length,  width  one-third  of  their 
length  ;  singly,  in  pairs,  and  long 
wavy  filaments. 

Colonies  circular  ;  at  first  trans- 
parent droplets,  later  yellowish  in 
colour. 

On  the  surface  of  gelatine  they 
form  a  greenish-yellow  film. 

In  broth  they  produce  turbidity, 
and  later  a  skin  on  the  surface,  and 
on  sterilised  fish  a  pale-yellow  sticky 
growth. 

Cultures  are  photogenic. 

They  were  isolated  from  the  sea 
at  Sydney. 

No.  II. — Rods  with  rounded  ends 
•27  /A  in  length,  '67  /x  in  width,  and 
filaments. 

Colonies  on  gelatine  are  circular 
with  sharp  contours  and  greyish- 
yellow  in  colour  ;  later  they  are 
irregular  and  granular. 

Inoculated  in  gelatine  the  bacilli 
form  a  greyish-white  filament  in 
the  track  of  the  needle,  and  a  shining 
patch  on  the  surface. 

On  the  surface  of  obliquely  solidi- 
fied gelatine  they  form  a  bluish- 
grey  film. 

In  broth  they  produce  only 
turbidity. 


Cultures  are  photogenic. 

They  were  isolated  from  phospho- 
rescent fish. 

No.  III. — Rods  not  so  thick  as 
those  of  No.  II.,  singly,  in  pairs,  and 
filaments.  They  are  motile. 

Colonies  are  white,  scaly,  and 
wrinkled. 

On  the  surface  of  gelatine  the 
growth  spreads  over  the  medium. 

On  agar  the  growth  is  scanty. 

In  broth  they  produce  turbidity 
and  a  skin  floating  on  the  surface. 

Cultures  are  photogenic  after  a 
few  days'  growth. 

They  were  isolated  from  a  piece 
of  cuttle-fish. 

Bacillus  argenteo-phosphores- 
cens  liquefaciens.— Rods  straight 
or  slightly  bent,  2  ^  in  length,  and 
in  width  one-third  of  their  length  ; 
filaments. 

Colonies  circular,  pale  brown  or 
pale  yellow  and,  after  liquefaction, 
with  radiating  processes  extending 
into  the  surrounding  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  growth  in  the  track 
of  the  needle,  and  near  the  surface 
a  cup-shaped  area  of  liquefaction. 

In  broth  they  produce  turbidity, 
and  form  a  skin  on  the  surface. 

On  sterilised  fish  they  form  a 
yellow  layer. 

They  are  photogenic  but  not 
markedly  so. 

Bacillus  aurantiacus  (Frank- 
land). — Rods  short  and  thick,  singly, 
in  pairs,  and  in  filaments. 

Colonies  are  prominent  and  pale 
orange  in  colour. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  slight  growth  in  the 
track  of  the  needle  and  an  orange 
patch  on  the  free  surface. 

On  agar  and  potato  the  growth 
is  also  orange. 

They  occur  in  water. 

Bacillus  aureus  (Adametz).— 
Slender  rods  straight  or  slightly 
bent,  1'5  to  4  p.  in  length,  and  '5  p. 
in  width  ;  in  pairs,  filaments,  and 
masses.  They  are  motile. 

Colonies  circular  or  oval  and 
yellow  in  colour. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  very  limited  in 


DESCRIPTION   OF   SPECIES. 


the  track  of  the  needle,  while 
small  chrome-yellow  hemispherical 
masses  develop  on  the  free  surface. 

On  potato  they  form  a  chrome- 
yellow  growth. 

They  occur  in  water  and  on  the 
skin. 

Bacillus  berolinensis  Indicus 
(Classen). — Slender  rods,  singly,  in 
pairs,  and  short  chains  ;  capsulated. 

Colonies  at  first  whitish  acquire 
in  a  few  days  an  indigo-blue  colour. 

On  the  surface  of  gelatine  they 
form  a  blue  layer,  which  slowly 
spreads. 

On  the  surface  of  agar  the  indigo- 
blue  colour  is  very  marked. 

On  potato  they  grow  abundantly, 
and  develop  the  same  colour.  The 
pigment  is  insoluble  in  alcohol, 
chloroform  or  water,  soluble  in 
strong  acids,  and  decolorised  by 
ammonia. 

They  were  isolated  from  river 
water  at  Berlin. 

Bacillus  brassicae  (Pommer).— 
Rods  1-9  to  5-4  n  in  length,  -91  to 
1'2  /LI  in  width,  and  filaments.  They 
form  spores. 

Colonies  have  the  appearance  of 
a  fine  mycelium. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  in  the  track  of  the 
needle  sends  off  fine  filaments,  and 
liquefaction  quickly  follows. 

In  the  depth  of  agar  white 
colonies  form  in  the  track  of  the 
needle,  and  on  the  surface  the 
growth  is  first  cloudy  and  later 
yellowish. 

They  were  isolated  from  infusion 
of  cabbage. 

Bacillus  brevis  (Mori).— Rods 
•25  p.  long  and  -8  p  broad.  Non- 
motile. 

Colonies  pale-yellow  ;  non-lique- 
fying. 

Inoculated  in  the  depth  of  gela- 
tine small  dots  appear  along  the 
needle  track  and  a  pale  yellowish 
growth  on  the  surface. 

On  agar  at  35°  C.  a  yellowish 
and  on  blood  serum  a  greyish 
growth  appears  in  two  or  three 
days.  They  do  not  grow  on  potato. 
In  broth  they  form  a  white  cloudy 
-deposit. 


Mice  inoculated  subcutaneously 
die  in  from  sixteen  to  thirty  hours. 
They  are  also  pathogenic  in  guinea- 
pigs  and  rabbits. 

They  were  found  in  drain  water. 

Bacillus  brunneus  (Adametz 
and  Wichman). — Rods  small  and 
slender.  Spore  formation  present. 

Colonies  at  first  white,  later 
brownish. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  occurs  along  the 
track  of  the  needle  and  also  on 
the  surface,  developing  a  brownish 
colour  in  the  surrounding  gelatine. 

They  occur  in  water. 

Bacillus  buccalis  fortuitus 
(Vignal).— Rods  1-4  to  3  /u  in 
length,  singly,  and  in  pairs. 

Colonies  circular  and  liquefying. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  slowly,  and 
white  flocculi  occur  in  the  liquid, 
and  later  subside  to  the  bottom. 

In  broth  they  produce  turbidity 
and  a  skin  on  the  surface. 

They  were  isolated  from  saliva. 

Bacillus  buccalis  maximus 
(Miller).— Rods  2  to  10  p  in  length, 
1  to  1-3  fjt.  in  width,  and  filaments 
30  to  150  /n  in  length. 

They  occur  in  the  mouth. 

Bacillus  buccalis  minutus  (Vig- 
nal).— Rods  -5  to  1  /ji.  in  length, 
and  slightly  less  in  width. 

Colonies  circular  and  faintly 
yellow. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  yellowish-white 
growth  in  the  track  of  the  needle, 
and  a  patch  of  the  same  colour  on 
the  surface  ;  liquefaction  com- 
mences slowly,  and  extends  down- 
wards until  the  gelatine  is  com- 
pletely liquefied  and  a  yellow  mass 
collects  at  the  bottom  of  the  tube. 
In  broth  there  is  a  similar  deposit, 
and  an  iridescent  pellicle  on  the 
surface. 

On  potato  they  form  a  yellow  film. 

They  were  isolated  from  saliva. 

Bacillus  butyricus,  (Praz- 
mowski.  Bacilhi,*  amylobacter,  Van 
Tieghem  ;  Bacillus  of  butyric  acid 
fermentation}. — Rods  3  to  10  ft  long, 
under  1  /*  wide,  often  indistinguish- 
able from  Bacillus  subtilis.  They 


DESCRIPTION   OF   SPECIES. 


503 


grow  out  into  long,  apparently  un- 
jointed  threads.  They  are  mostly 
actively  motile,  but  also  occur  in 
zoogloea.  The  rods  and  threads 
are  sometimes  slightly  bent,  like 
vibrios.  They  are  anaerobic.  The 
shorter  rods  as  a  rule  swell  in  the 
middle,  becoming  ellipsoidal,  lemon, 


FIG.  199.— CLOSTRIDIUM  BUTTKICUM. 

A.  Active  stage,  (a,  6)  Bent  rods  (vibrio- 

form)    and    threads.      (c)    Short 
rods,     (d)  Long  rods. 

B.  Spore-formation.    C.  Spore-germina- 

tion.    (Prazmowski. ) 

or  spindle-shaped  ;  the   long  rods,  ; 

and  sometimes  the  short  ones,  swell  ! 

at  one  end  ;  in  either  case  ellipsoidal  ! 
spores  are  developed  (Fig.  199). 

Cultivated  in   nutrient   gelatine,  j 

the  medium  is  liquefied,  and  a  scum  j 

formed  on  the  surface.     They  grow  [ 


best  between  35°  and  40°  C.  The 
spores  are  widely  distributed  in 
nature,  and  grow  readily  on  fleshy 
roots,  old  cheese,  etc.  They  convert 
the  lactic  acid  in  milk  into  butyric 
acid,  and  produce  the  ripening  of 
!  cheese. 

They  occur  also  in  solutions  of 
starch,  dextrine,  and  sugar,  and  are 
the  active  agents  in  the  fermenta- 
tion of  sauerkraut  and  sour  gherkins. 
Bacillus  foutyricus  (Botkin).— 
Rods  and  filaments,  spore -formation 
present.  They  are  anaerobic. 

Colonies  consist  of  felt-like 
masses. 

Inoculated  in  the  depth  of  gela- 
tine with  1-5  per  cent,  of  grape- 
sugar,  the  growth  commences  in 
the  lower  part  of  the  needle  track 
with  abundant  formation  of  gas 
bubbles  and  liquefaction  of  the 
jelly. 

In  milk  there  is  abundant  gas 
formation,  which  will  break  the 
flasks  if  closed. 

They  were  isolated  from  milk, 
earth,  and  water. 

Bacillus        butyricus 
n    (Hueppe). — Rods  slightly  bent, 
*     2-1  p  in  length,  -38  /*  in  width, 
and  filaments. 

Colonies  yellowish ;  rapidly 
liquefying. 

Inoculated  in  the  depth  of 
gelatine  liquefaction  occurs 
along  the  track  of  the  needle,  and 
later  a  wrinkled  skin  floats  on 
the  surface. 

On  agar  the  growth  is  yel- 
lowish. 

On  potato  the  growth  is 
wrinkled  and  faintly  yellow. 

They  coagulate  milk,  precipita  • 
ting  and  then  dissolving  the  casern. 
They  occur  in  milk. 
Bacillus  cadaveris  (Sternberg). 
— Rods   1 '5  to  4  /A  in  length,  and 
1*2  /LI  in  width,  singly,  in  pairs,  and 
in  filaments. 

They  are  anaerobic.  Colonies  in 
glycerine-agar  are  irregular,  granu- 
lar, and  white. 

They  produce  an  acid  reaction 
in  cultures. 

Subcutaneous  injection  in  guinea- 
pigs  may  produce  extensive 


504 


DESCRIPTION   OF   SPECIES. 


oedema  and  death  in  twenty-four 
hours. 

They  were  obtained  from  the 
liver  in  fatal  cases  of  yellow  fever. 

Bacillus  caeruleus  (Smith).— 
Rods  2  to  2-5  \i  in  length,  and  '5  /* 
in  width,  singly  and  in  chains. 

Colonies  blue. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  colourless  growth 
in  the  track  of  the  needle,  and  a 
cup-shaped  cavity  in  its  upper  part, 
with  bluish  contents. 

On  agar  they  form  a  blue  layer 
and  a  deep-blue  growth  on  potato. 

They  occur  in  water. 

Bacillus     canalis     capsulatus 
(Mori).— Rods  '9  to  1*6  p.  in  width,  i 
capsulated. 

Colonies  milk-white. 

The  growth  in  the  depth  of  gela- 
tine is  similar  to  Friedlander's 
pneumococcus. 

On  agar  the  growth  is  viscid,  and 
on  potato  it  is  yellowish. 

In  broth  a  skin  forms  on  the 
surface. 

They  are  pathogenic  in  mice. 

They  occur  in  sewage. 

Bacillus  canalis parvus  (Mori). 
-Rods  2  to  5  p.  in  length,  -8  to  1  p 
in  width. 

Colonies  very  minute,  pale  yellow. 

On  the  surface  of  gelatine  they 
very  slowly  form  a  yellowish  film. 

On  agar  the  growth  is  dry  and 
yellow. 

They  are  pathogenic  in  mice  and 
guinea-pigs. 

They  occur  in  sewage. 

Bacillus  candicans  (Frankland). 
— Very  short  rods  and  filaments. 

Colonies  pure  white. 

Inoculated  in  the  depth  of  gela- 
tine they  form  isolated  colonies  in 
the  track  of  the  needle,  and  a  white 
button  on  the  free  surface. 

On  agar  they  form  a  greyish 
layer,  and  flourish  on  potato. 

They  occur  in  soil. 

Bacillus  capsulatus  (Mori). — 
Oval  forms  and  rods,  sometimes 
encapsuled.  Non-motile.  Colonies 
white. 

Inoculated  in  the  depth  of  gela- 
tine and  agar  a  nail-shaped  growth 
occurs. 


In  broth  they  form  a  white- 
turbidity,  and  a  white  pellicle 
develops  on  the  surface  and  on  the 
sides  of  the  vessels. 

On  potato  at  37°  C.  an  abundant 
moist,  yellowish,  stringy  growth  is 
formed,  with  production  of  gas 
bubbles.  They  are  pathogenic  in 
mice  and  in  rabbits  if  injected  into 
the  pleural  cavity. 

They  occur  in  drain  water. 

Bacillus  capsulatus  (Pfeiffer). 
- — Rods  singly,  in  pairs,  or  in  chains 
and  in  filaments.  They  have  a  well- 
marked  capsule.  Colonies  white. 

Inoculated  in  gelatine  they  grow 
in  the  track  of  the  needle,  and 
form  a  white  button  on  the  free 
surface. 

On  agar  and  on  potato  the  growth 
is  also  white  and  very  viscid,  so 
that  it  can  be  drawn  out  into  long 
threads. 

They  produce  a  fatal  result  in 
mice  in  two  or  three  days,  when 
inoculated  subcutaneously.  A 
minute  quantity  of  a  broth  culture 
injected  into  the  peritoneal  cavity 
of  guinea-pigs  will  prove  fatal  in 
thirty-six  hours.  The  bacilli  are 
found  in  the  blood  which  is  made 
viscid. 

They  were  isolated  from  a  guinea- 
pig  found  dead. 

Bacillus  capsulatus  mucosits 
(Fasching). — Rods  3  to  4  p  in 
length,  '75  to  1  ju,  in  width  ;  capsu- 
lated. Colonies  white. 

Cultures  in  gelatine  resemble 
Friedlander's  pneumococcus. 

They  form  gas. 

They  produce  a  fatal  result  in 
mice  in  thirty- six  hours. 

They  wrere  isolated  from  nasal 
mucus  in  cases  of  influenza. 

Bacillus  capsulatus  suis 
(Smith).— Rods  from  1-2  to  1'8  /z 
in  length  and  '8  to  '9  /j.  in  width. 
There  are  three  varieties  of  this 
bacillus  having  a  close  resemblance  to 
the  pneumococcus  of  Friedliinder. 

They  were  isolated  from  the  in- 
testines of  swine. 

Bacillus  carabiformis  (Kac- 
zynsky). — Rods  short  and  slender. 

Colonies  develop  characteristic 
processes. 


DESCRIPTION    OF   SPECIES. 


505 


Inoculated  in  the  depth  of  gela- 
tine, the  bacilli  produce  liquefaction 
and  colour  the  liquid  greenish- 
yellow. 

On  agar  they  form  a  yellowish- 
white  layer. 

They  were  isolated  from  the 
stomach  of  a  dog. 

Bacillus  carnicolor  (Tils).— 
Rods  2  ft  long,  and  *5  ft  broad. 
Singly  actively  motile.  Spore- 
formation  not  observed. 

Colonies  are  in  the  form  of  cup- 
shaped  depressions. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  rapidly  along  the 
whole  track  of  the  needle,  forming 
a  funnel-shaped  area  of  liquefaction 
at  the  bottom  of  which  there  is  a 
pale  pink  deposit. 

On  potato  they  form  slowly  a 
dark  flesh-coloured  growth. 

They  occur  in  water. 

Bacillus  carotarum  (A.  Koch). 
—Rods  -97  to  1'05  p.  in  length  and 
filaments. 

Colonies  white  and  circular. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  grow  slightly  in  the 
track  of  the  needle  and  abundantly 
on  the  surface. 

On  agar  they  form  a  white,  and 
on  potato  a  faintly  brown  layer. 

They  occur  on  boiled  carrot  and 
beet. 

Bacillus  cavicida  (Brieger). — 
Rods  morphologically  and  in  culti- 
vations similar  to  Bacillus  coli  com- 
munis. 

Cultures  are  said  to  be  patho- 
genic in  guinea-pigs. 

They  were  isolated  from  fseces. 

Bacillus  cavicida  Havaniensis 
(Sternberg). — Rods  2  to  3  ft  in 
length,  and  •?  ft  in  width,  singly 
and  in  pairs. 

Colonies  are  of  a  pale-straw 
colour. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  form  small  trans- 
lucent pearl-like  spherical  colonies, 
and  on  the  free  surface  the  growth 
is  limited. 

On  potato  the  growth  is  at  first 
thin  and  dirty  yellow,  and  later 
gamboge  yellow. 

Guinea-pigs  inoculated  subcuta- 


neously  die  in  ten  or  twelve 
hours. 

They  were  isolated  from  the 
intestinal  contents  in  a  fatal  case 
of  yellow  fever,  by  inoculation  of 
guinea-pigs. 

Bacillus  chromo-aromaticus.— 

Rods  which  liquefy  gelatine  and 
form  a  yellowish-white  scum  on 
the  surface. 

On  potato  the  growth  is  irides 
cent  and  brownish. 

In  broth  a  scum  forms  on  the 
surface  and  the  broth  is  coloured 
greenish-blue.  Cultures  have  an 
aromatic  odour. 

They  are  said  to  produce  pneu- 
monia and  pleurisy  in  rabbits. 

They  were  isolated  from  a  pig 
with  postmortem  appearances  of 
swine-fever. 

Bacillus  circulans  (Jordan). — 
Rods  2  to  5  fi  in  length  and  1  ft  in 
width,  singly  and  in  short  chains. 

Colonies  are  brownish. 

Inoculated  in  the  depth  of  gela- 
tine they  liquefy  the  medium  in 
the  track  of  the  needle,  forming 
a  conical  cavity  at  its  upper  part. 

On  agar  they  form  a  translucent 
film. 

Milk  is  slowly  coagulated. 

In  broth  they  produce  turbidity 
and  a  slimy  deposit. 

They  occur  in  water. 

Bacillus  citreus  cadaveris 
(Strassmann). — Rods  '9  ft  in  length, 
•6  ft  in  width,  singly  and  in  chains. 

Colonies  pale  yellow. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  form  minute  colonies 
along  the  track  of  the  needle,  and 
at  its  upper  part  liquefy  the  gela- 
tine and  produce  a  yellow  deposit. 

They  were  found  in  the  blood 
after  death. 

Bacillus  cloacae  (Jordan).— 
Short  rods  '8  to  1-9  ft  in  length, 
•7  to  1  ^  in  width,  singly  and  in 
pairs. 

Colonies  circular,  yellowish. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the  track 
of  the  needle,  an  iridescent  scum 
forms  on  the  surface,  and  there  is. 
an  abundant  deposit. 

On    agar   the    growth    is    milk- 


306 


DESCRIPTION   OF   SPECIES. 


white,  and  on  potato  yellowish- 
white. 

In  broth  they  produce  turbidity 
and  a  scum  on  the  surface.  They 
reduce  nitrates. 

They  occur  in  sewage. 

Bacillus  coli  communis 
(Escherich).— See  p.  344. 

Bacillus  coli  similis  (Stern- 
berg). — Rods  1  to  3  /A  in  length,  -4  to 
•5  p.  in  width  ;  singly  and  inpairs. 

Colonies  circular  and  pale  brown 
in  colour. 

In  the  depth  of  gelatine  they 
form  a  scanty  growth  in  the  track 
of  the  needle,  and  on  the  free  sur- 
face a  translucent  film  with  irregu- 
lar margins. 

On  potato  the  growth  is  pale 
brown  or  dirty  white. 

They  were  isolated  from  human 
diver  after  death. 

Bacillus  constrictus  (Zimmer- 
mann). — Rods  from  1'5  to  6'5  p.  in 
length,  and  -75  /u,  in  width.  The 
rods  are  segmented. 

Colonies  are  circular,  granular, 
and  greyish-yellow. 

In  the  depth  of  gelatine  they 
form  a  filament  in  the  track  of  the 
needle  and  irregular  yellow  heaps 
on  the  free  surface. 

On  the  surface  of  agar  the  growth 
consists  of  a  yellow  shinin^layer, 
and  on  potato  the  same  colour  is 
produced. 

They  occur  in  water. 

Bacillus  coprogenes  foetidus 
(Schottelius). — Rods  about  as  large 
as  Bacillus  subtilis,  but  shorter. 
They  are  non-motile.  Spore-for- 
mation occurs  when  the  bacilli  have 
access  to  the  air,  but  not  in  the 
-animal  body. 

In  the  depth  of  gelatine  a  fila- 
ment forms  composed  of  yellow- 
ish compact  colonies  ;  and  on  the 
surface  a  fine  transparent  film ; 
cultures  emit  a  strong  putrefactive 
odour. 

On  potato  they  form  a  light  grey, 
dry  layer. 

Subcutaneous  injection  of  small 
•doses  had  no  effect  on  mice  and 
rabbits,  but  very  large  quantities 
produced  a  toxic  effect  in  rabbits. 
Swine  are  not  affected. 


They  were  found  by  Schottelius 
in  the  intestine  in  cases  of  swine 
erysipelas. 

Bacillus  coprogenes  parvus 
(Bienstock). — Very  short  rods. 

On  the  surface  of  gelatine  they 
form  a  very  limited,  almost  in- 
visible, growth  in  the  track  of  the 
needle. 

In  mice  they  produce  oedema  and 
J  death  in  thirty-six  hours,  and  in 
|  rabbits  a  local  rash  and  death  in 
eight  days. 

They  were  isolated  from  human 
evacuations. 

Bacillus  crassus  aromaticus 
(Taratoff). — Rods  3'5  to  5  p  long, 
1-5  \i  in  width  ;  constricted  in  the 
centre. 

Colonies  appear  in  the  form  of 
cup-shaped  depressions  and  pro- 
duce a  fruit-like  odour. 

In  gelatine  they  grow  in  the 
track  of  the  needle,  and  later 
produce  a  funnel-shaped  area  of 
liquefaction. 

They  occur  in  well  water. 

Bacillus  crassus  sputigenus 
(Kreibohm). — Short  thick  rods, 
sometimes  curved;  capsulated. 
Colonies  greyish- white.  Cultures 
in  gelatine  resemble  those  of  Fried- 
lander's  pneumococcus.  They  are 
pathogenic  in  small  animals.  They 
were  isolated  from  human  sputum. 

Bacillus  cuniculicida,  Bacillus 
of  Rabbit  Styticcemia  (see  p.  228). 

Bacillus  cuticularis  (Tils).— 
Rods  from  2  to  3  p.  in  length,  '3  to 
"5  ft  in  width,  and  filaments. 

Colonies  are  yellow  and  the  gela- 
tine is  liquefied. 

In  the  depth  of  gelatine  they 
produce  liquefaction,  and  a  skin 
forms  on  the  surface. 

On  potato  the  growth  is  slimy 
and  yellow. 

They  occur  in  water. 

Bacillus  cuticularis  a  1  b  u  s 
(Taratoff).— Rods  3'2  p  long,  con- 
stricted in  the  middle.  Actively 
motile. 

Colonies  are  opalescent  and 
bluish-white. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  white  rosette- 
shaped  growth  on  the  surface  and 


DESCRIPTION   OF   SPECIES. 


507 


a  shining  white  growth 
along  the  needle  track 
which  sends  off  long 
rounded  processes. 

On  agar,  glycerine  agar 
and  blood  serum  they 
produce  a  luxuriant  white 
shining  growth. 

Broth  becomes  turbid 
with  a  whitish  deposit  and 
pellicle. 

On  potato  there  is  a  thick 
moist  brown  growth. 

They  are  found  in  water. 

Bacillus  cyaneo-fuscus 
( Bey erinck).— Rods  '2  to  -6 
JJL  in  length,  and  half  their 
length  in  width.  Motile. 

In  the  depth  of  gelatine 
they  produce  colonies  in 
the  track  of  the  needle, 
which  later  develop  a  black 
pigment. 

In  broth  with  ^  per  cent, 
of  peptone  they  produce  a 
blue  colour,  changing  to 
brown  and  finally  black. 

They  were  isolated  from 
cheese,  size  and  glue. 

Bacillus  cyaneo-phosphores- 
cens  (Katz).— Rods  2'6  p.  in  length, 
1  n  in  width,  singly,  in  pairs,  and 
.filaments.  Colonies  circular  and 
brownish  or  greyish-yellow. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  grey-white  fila- 
ment in  the  track  of  the  needle  and 
liquefaction  follows  at  the  upper 
part  ;  later  a  skin  forms  on  the  sur- 
face and  a  yellow  deposit  occurs  at 
the  bottom  of  the  liquefied  jelly 
which  has  a  reddish-brown  tinge. 

In  broth  a  similar  skin  floats 
on  the  surface  and  the  broth  is 
turbid. 

On  sterilised  fish  the  growth  is 
viscid  and  yellow. 

Cultures  are  phosphorescent, 
especially  in  media  containing 
excess  of  common  salt. 

They  were  isolated  from  sea- 
water  at  Sydney,  and  are  possibly 
identical  with  Bacillus  phosphores- 
cens  of  Fischer. 

Bacillus  cyanogenus  (Hueppe). 
— Bacterium  syncijiinum.  liari/l//* 
•of  Blue  Milk.—  Motile  rods,  2'5  to 


c      . 
•••»,«?•* 


\\ 


/\, 


;\ 


FIG.  200. — BACILLUS  CYANOGENUS,  x  650.  A.  Ac- 
tive rods.  B  Rods  in  zooglpea.  C.  Chain  of 
short  rods.  D.  Chain  of  cocci.  E.  Cocci  stage. 
F,  G.  Spore-forming  rods.  H.  Involution- 
forms  (NEELSEN). 


3-5  n  in  length,  and  -4  n  wide  (Fig. 
200).  The  rods  after  division  may 
remain  linked  together,  and  form 
chains.  Non-motile  rods  occur 
enveloped  in  a  gelatinous  capsule, 
and  involution  forms. 

Colonies  appear  after  two  days 
as  small  greyish-white  points  which 
gradually  assume  a  moist  appear- 
ance. The  gelatine  becomes  steel- 
grey,  throwing  the  white  colonies 
into  strong  relief. 

In  the  depth  of  gelatine  a  whitish 
growth  appears  in  the  track  of  the 
needle  near  the  upper  part,  and  on 
the  free  surface,  producing  also  a 
dark  steel-blue  discoloration  of 
the  jelly  which  spreads  down- 
wards. 

On  agar  a  greyish  growth  ap- 
pears, and  the  agar  is  coloured  dark 
brown. 

On  potato  a  yellowish  moist 
growth  develops,  the  potato  around 
it  is  stained  grey-blue.  Milk  be- 
comes slightly  alkaline  and  of  a 
slate-grey  colour,  which  on  the 
addition  of  acid  changes  to  an 
intense  blue.  Milk  in  which  the 


508 


DESCRIPTION    OF   SPECIES. 


lactic  acid  bacillus  is  growing  be- 
comes sky-blue  from  the  first. 

They  are  non-pathogenic. 

They  are  present  in  blue  milk. 

Bacillus  cyanogenus  (Jordan). 
— Rods  1-3  p.  in  length,  '8  p.  in 
width.  Slightly  motile. 

Colonies  granular  and  irregular. 
They  colour  the  surrounding  gela- 
tine brown. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  produce  a  scanty 
growth  in  the  track  of  the  needle, 
and  a  film  on  the  surface  with 
coloration  of  the  gelatine  beneath 
it. 

On  agar  they  form  a  white  layer, 
and  the  jelly  is  coloured  brown. 

On  potato  the  growth  is  brown. 

They  were  isolated  from  sewage. 

Bacillus  cystiformis  (Clado). — 
Short  slender  rods.  Motile. 

Colonies  circular,  yellowish,  gra- 
nular. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  scanty  growth  in  the 
track  of  the  needle,  and  a  white 
patch  on  the  free  surface. 

On  agar  the  growth  is  yellowish- 
white. 

They  were  isolated  from  urine. 

Bacillus  delicatulus  (Jordan), 
r— Rods  2  p,  long  and  1  p,  broad, 
often  in  pairs  or  short  chains. 
Actively  motile.  Spore-formation 
not  observed. 

Colonies  at  first  whitish  with 
a  radiating  edge.  Later  they 
liquefy  the  gelatine  and  the  centres 
become  dark. 

Inoculated  in  the  depth  of  gela- 
tine they  rapidly  liquefy  it  and 
form  a  whitish  pellicle  and  a  brown 
deposit. 

On  agar  a  greyish  crinkled 
growth  appears,  which  gradually 
becomes  white  and  shining. 

On  potato  there  is  a  grey  flat 
growth. 

Milk  is  coagulated  and  becomes 
strongly  acid. 

Broth  is  made  turbid  and  a  white 
serum  and  precipitate  formed. 

They  occur  in  sewage. 

Bacillus  dentalis  viridans 
(Miller).— Rods  slightly  bent,  singly 
and  in  pairs. 


Colonies  circular,  yellowish  and 
concentric. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  in  the  track  of  the 
needle  and  on  the  free  surface,  and 
the  jelly  is  coloured  green. 

On  agar  the  growth  is  colourless 
or  slightly  grey. 

Intraperitoneal  injections  in  mice 
and  guinea-pigs  produce  a  fatal  re- 
sult. Subcutaneous  injections  cause 
suppuration. 

They  were  isolated  from  caries 
of  the  teeth. 

Bacillus  dentriticus  (Bordoni 
Uffreduzzi  and  Lustig).  —  Rods  "85 
to  2-8  p.  long,  and  '5  to  -85  p.  broad, 
singly  and  in  zoogloea.  Motile. 
Spore-formation  not  observed. 

Colonies  have  an  arborescent 
appearance. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  circular  raised 
growth  at  the  point  of  puncture, 
and  white  colonies  along  the  needle 
track.  The  jelly  is  gradually 
liquefied. 

On  agar  and  blood  serum  there 
is  a  scanty  growth  on  the  surface 
and  an  abundant  growth  in  the 
track  of  the  needle.  Blood  serum 
is  liquefied  after  some  time. 

Broth  is  rendered  turbid  ;  a 
white  firm  pellicle  forms. 

On  potato  there  is  a  thick  moist 
white  growth,  which  later  becomes 
yellow. 

Found  in  water. 

Bacillus  devorans  (Zimmer- 
mann).  —  Rods  -99  p,  to  1*2  p.  in 
length,  '74  p.  in  width  ;  singly,  in 
pairs  and  in  chains. 

Colonies  are  circular,  granular, 
and  grey,  with  periphery  formed  of 
radiating  processes. 

In  the  depth  of  gelatine  they 
produce  a  whitish  filament  and  an 
excavation  at  the  upper  part,  which 
may  or  may  not  contain  liquefied 


On  agar  a  greyish  film  is  found. 

They  do  not  grow  on  potato. 

They  occur  in  water. 

Bacillus  diffusus  (Frankland). 
—  Rods  1/7  p.  in  length,  '5  p.  in 
width  ;  singly,  in  pairs,  and  fila- 
ments. 


DESCRIPTION   OF   SPECIES. 


509 


Colonies  circular,  bluish-green, 
•with  a  granular  nucleus  and  delicate 
irregular  periphery. 

In  the  depth  of  gelatine  there  is 
scarcely  any  growth  in  the  track  of 
the  needle,  but  a  shining  greenish- 
yellow  film  on  the  surface,  and 
liquefaction  below  it. 

On  agar  the  growth  is  faintly 
yellow. 

In  broth  they  produce  turbidity 
and  a  yellowish  deposit. 

On  potato  the  growth  is  yellow- 
ish. 

They  occur  in  earth. 

Bacillus  diphtheriae  (p.  332). 

Bacillus  diphtheriae  colum- 
barum  (p.  336). 

Bacillus  dysodes(Zopf).— Cocci, 
long  and  short  rods,  and  spores. 

They  were  observed  in  bread, 
making  it  greasy  and  unfit  for  food, 
and  generating  a  penetrating  odour 
resembling  a  mixture  of  pepper- 
mint and  turpentine.  A  great  loss 
may  result  to  bakers  if  the  fungus 
is  introduced  with  the  yeast. 

Bacillus  endocarditidis  cap- 
sulatus  (Weichselbaum).— Cocci 
resembling  Fried  lander's  pneumo- 
cocci. 

Colonies  faintly  yellow,  with 
dentated  contours. 

In  the  depth  of  gelatine  the 
growth  produces  a  filament  in  the 
track  of  the  needle,  and  a  patch  like 
stearin  on  the  free  surface. 

Large  doses  injected  subcutane- 
ously,  or  into  the  peritoneal  cavity 
prove  fatal  to  rabbits. 

They  were  isolated  from  infarcts 
in  a  fatal  case  of  endocarditis. 

Bacillus  endocarditidis  griseus. 
—Rods  motile. 

Colonies  granular  and  brown  or 
yellowish-brown. 

In  the  depth  of  gelatine  there  is 
a  filamentous  growth  in  the  track 
of  the  needle,  and  a  circular  whit- 
ish patch  on  the  surface.  On  agar 
and  potato  the  growth  is  greyish- 
brown. 

Cultures  cause  a  fatal  result  in 
mice  and  guinea-pigs. 

They  were  isolated  from  a  case  of 
endocarditis. 

Bacillus  enteritidis  (p.  372). 


Bacillus  epidermidis  (Bordoni 
Uffreduzzi).— Rods  2-8  to  3  /i  in 
length,  and  -3  \i  in  breadth.  Spore- 
formation  occurs  at  25°  C. 

They  grow  very  sparingly  on 
gelatine. 

On  agar  there  is  a  surface  growth. 

On  potato  at  15°  to  20°  C.  the 
growth  appears  first  in  the  form 
of  drops,  which  gradually  extend 
and  coalesce  and  form  a  thin  layer 


i-NN . 


!  FIG.  201. — PURE  CULTIVATION  OF  BA- 
CILLUS FHJUKAXS  ON  THE  SURFACE 
OF  NUTRIENT  AGAR-AGAR. 

I  over  the  surface.     On  blood  serum 
they  form  a  thin  film. 

Inoculation  in  rabbits  and  guinea- 
i  pigs  and  on  the  human  skin  pro- 
|  duced  no  result. 

They  were  isolated  from  flakes 
of  cuticle  from  between  the  toes. 

Bacillus  erysipelatis  suis  (p. 
356). 

Bacillus  erythrosporus 
i  (Eidam).  —  Slender  rods  and 


510 


DESCRIPTION    OF   SPECIES. 


filaments.      Motile, 
tion  present. 


Spore-forma-  I       On  potato  the  growth  is  brown. 
They  occur  in  water. 


FIG.  202.— PHOTOGRAPH  OF  PART  OF  AN  IMPRESSION  PREPARATION  OF 
BACILLUS  FIGURANS  ON  NUTRIENT  GELATINE,  x  50. 


Colonies  circular,  with  brown  nu- 
cleus and  yellowish-green  periphery. 
In   the   depth   of   gelatine   they 


Bacillus  figurans  (Crookshank). 
— Rods,  with  rounded  ends,  varying 
in  length.  Spore-formation  present. 


FIG.  203.— PART  OF  THE  SAME  SPECIMEN  SHOWN  IN  FIG.  202 


x  200. 


grow  in  the  track  of  the  needle 
and  on  the  free  surface,  colouring 
the  jelly  green  by  transmitted  light, 
yellow  by  reflected  light. 


In  plate-cultivations  they  cause 
a  cloudy  growth,  spreading  from 
various  points ;  if  a  cover-glass 
impression  is  made,  this  is  found 


DESCRIFFION   OF   SPECIES. 


511 


to  consist  of  the  regularly-arranged 
parallel  rods.  The  chains  of  rods 
become  twisted  at  intervals  into 
curious  convolutions,  from  which 
offshoots  are  continued  in  various 
directions.  These  long  shoots  or 
processes  are  again  twisted  at 
intervals  into  varying  shapes  and 
patterns  (Figs.  202,  203).  Culti- 
vated in  nutrient  gelatine,  the  bacilli 
form  on  the  surface  visible  windings, 
from  which  fine  filaments  grow 
down  into  the  gelatine.  They 
spread  out  also  in  almost  parallel 
lines  transversely  from  the  needle 
track.  The  gelatine  is  not  liquefied. 

On  an  oblique  surface  of  nutrient 
agar-agar  the  filaments  spread  down- 
wards into  the  substance  of  the 
jelly,  and  outwards  from  the  cen- 
tral streak  on  the  surface,  forming 
a  feather-like  cultivation  (Fig.  201). 

They  were  obtained  from  the 
air,  and  later  were  identified  by 
the  author  with  Bacterium  Zopfii. 

Bacillus  figurans  (Vaughan).— 
Rods  and  threads. 

Colonies  composed  of  curved  and 

terlacing  lines. 

In  gelatine  they  grow  in  the  track 
of  the  needle,  and  very  slowly 
liquefy  it. 

On  agar  they  form  a  thin  white 
layer. 

They  were  found  in  water. 

Bacillus  filiformis  (Tils).— Rods 
4  p.  in  length  and  1  /z  in  width, 
singly  and  in  chains.  Spore-forma- 
tion present. 

Colonies  are  granular,  with  yel- 
lowish nucleus. 

Inoculated  in  the  depth  of  gela- 
tine there  is  no  growth  in  the 
track  of  the  needle,  but  a  whitish 
growth  on  the  surface,  liquefying 
the  jelly  slowly. 

In  broth  they  form  a  skin  on 
the  surface. 

On  agar  the  growth  is  white. 

On  potato  the  growth  is  dry  and 
after  a  time  brownish.  They 
coagulate  milk. 

They  occur  in  water. 

Bacillus  filiformis  Havaniensis 
(Sternberg). — Long  slender  rods, 
3  /A  in  diam.,  and  filaments. 

Colonies  circular,  irregular  ;  deep 


colonies  are  brownish ;  superficial 
colonies  thin  and  translucent. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  scanty  in  the 
track  of  the  needle. 

In  the  depth  of  agar  an  opaque 
branching  growth  occurs  in  the 
track  of  the  needle,  and  a  scanty 
milk-white  growth  on  the  surface. 

In  broth  they  cause  opalescence. 

They  were  isolated  from  the 
liver  in  fatal  cases  of  yellow  fever. 

Bacillus  flavescens  (Pohl).— 
Rods  2-1  to  2-2  fj,  in  length,  -8  /A 
in  width.  Slightly  motile. 

Colonies  yellow  and  granular. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  produce  a  filament 
in  the  track  of  the  needle,  and  a 
growth  spreads  over  the  free  sur- 
face. 

On  the  surface  of  agar  the  growth 
is  composed  of  isolated  yellow 
colonies. 

On  potato  they  grow  rapidly, 
forming  a  shiny  yellow  layer. 

They  occur  in  marsh  water. 

Bacillus  flavocoriaceus  (Ada- 
metz). — Minute  rods  occurring  in 
zoogloea. 

Colonies  circular,  sulphur-yellow. 
Under  a  low  power  they  show  a 
brownish-yellow  nucleus  and  yellow 
periphery. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  granular  in  the 
track  of  the  needle  and  on  the  free 
surface. 

They  occur  in  water. 

Bacillus  flue-res  cens  aureus 
(Zimmermann). — Short  rods,  1/9  \L 
in  length,  '74  p.  in  width. 

Singly,  in  pairs  and  in  masses. 
Motile  ;  flagellated. 

Coloniescircular,  granular,  yellow. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  filament  in  the 
track  of  the  needle  and  a  yellow 
patch  on  the  surface. 

On  agar  the  growth  is  golden- 
yellow,  and  the  same  on  potato. 

They  occur  in  water. 

Bacillus  fluorescens  lique- 
faciens  (Flugge). — Short  rods  with 
rounded  ends. 

Colonies  on  plates  develop  an 
iridescence  around  them. 


512 


DESCRIPTION   OF   SPECIES. 


In  the  depth  of  gelatine  a  white 
filament  forms  in  the  track  of  the 
needle,  with  liquefaction  in  the 
upper  part,  and  an  iridescent  sheen 
is  produced  in  the  jelly. 

On  potato  they  develop  a  brown- 
ish layer. 

They  occur  in  water  and  in 
putrid  infusions. 

Bacillus  fluorescens  lique- 
faciens  minutissimus  (Unna  and 
Tommasoli). — Rods  1  '5  to  2  /u.  in 
length,  -3  p.  in  width. 

Colonies  circular,  with  brownish 
nucleus  and  yellowish  marginal 
zone. 

In  the  depth  of  gelatine  lique- 
faction forms  in  the  track  of  the 
needle.  The  liquid  is  turbid  and 
yellowish,  and  a  white  sediment 
forms  at  the  bottom  and  a  fluores- 
cent scum  on  the  surface. 

On  agar  and  potato  the  growth  is 
brownish. 

They  were  isolated  from  eczema- 
tous  skin. 

Bacillus  fluorescens  longus 
{Zimmermann). — Rods  varying  in 
length,  some  curved,  1'45  to  1'65  p 
in  length,  '83  p,  in  width,  and  wavy 
filaments  14  p  in  length.  Motile. 

Colonies  circular,  well-defined, 
yellowish,  with  broad  twisted 
markings. 

On  the  surface  of  gelatine  they 
produce  a  layer  with  a  bluish-green 
fluorescent  colour. 

On  agar  a  thin  film  forms,  and 
the  jelly  is  coloured  greenish- 
yellow. 

On  potato  the  growth  is  slimy 
and  yellowish. 

They  occur  in  water. 

Bacillus  fluorescens  nivalis 
(Schmolck). — Rods  and  chains. 
Motile. 

Colonies  circular,  surrounded  by 
liquid  fluorescent  jelly. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  in 
the  track  of  the  needle,  and  the 
jelly  is  coloured  green. 

On  agar  the  growth  is  white  and 
the  jelly  green. 

On  potato  the  growth  is  brownish. 

They  are  probably  identical  with 
Bacillus  fluorescens  liquefaciens. 


They  were  isolated  from  snow. 

Bacillus  fluorescens  non-lique- 
faciens  (Eisenberg). — Short  deli- 
cate rods.  Non-motile. 

Colonies  have  the  appearance 
of  fern-leaves,  and  are  opales- 
cent. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  very  slight  in 
the  track  of  the  needle,  and  on  the 
surface  filmy  and  fluorescent. 

On  agar  they  form  a  greenish 
layer. 

They  produce  a  brownish  layer 
on  potato,  and  bluish-grey  dis- 
coloration. 

They  occur  in  water. 

Bacillus  fluorescens  putidus 
(Fltigge). — Short  rods  with  rounded 
ends.  Motile  ;  spore-formation  not 
known. 

They  form  small  dark  colonies 
with  a  greenish  sheen,  which  have 
a  penetrating  odour. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  a  pale-grey 
growth,  and  after  three  days  colour 
the  medium  with  a  greenish  tinge 
spreading  down  from  above. 

On  potato  they  rapidly  develop 
a  brownish  layer. 

They  occur  on  decomposing  sub- 
stances, producing  a  greenish 
coloration. 

According  to  Lehmann  and  Neu- 
mann the  Bacillus  fluorescens  albus 
and  Bacillus  fluorescens  longus  of 
Zimmermann  and  Bacillus  fluores- 
cens non-liquefaciens  are  merely 
varieties  of  Bacillus  fluorescens 
putidus  ;  and  further,  cultures  of 
this  bacillus  on  agar  and  on  potato 
and  in  milk  and  in  broth  cannot 
be  distinguished  from  those  of 
Bacillus  fluorescens  liquefaciens. 

Bacillus  fluorescens  tenuis 
(Zimmermann). — Rods  1  to  1'85  fj. 
in  length,  '8  p.  in  width.  Singly, 
in  masses,  and  filaments.  Motile. 

Colonies  irregular. 

Inoculated  in  the  depth  of  gela- 
tine a  delicate  filament  forms  in 
the  track  of  the  needle,  and  on 
the  surface  a  greyish-white  growth 
spreads  and  colours  the  gelatine 
yellow. 

On  agar  the  growth   is   shining 


DESCRIPTION   OF   SPECIES. 


513 


and  greenish,  and  on  potato  yellow- 
ish. 

They  occur  in  water. 

Bacillus  foetidus  (Bacterium 
fcetidum,  Thin). — Cocci,  short  rods, 
long  rods,  and  leptothrix.  The 
cocci,  1'25  to  1-4  /A  in  diam.,  occur 
singly  or  in  pairs.  Spore-formation 
present  in  the  rods. 

They  were  isolated  from  the 
exudation  in  a  case  of  profuse 
sweating  of  the  feet,  and  the  odour 
was  noticeable  in  the  cultivation 
Pi<i<;Ulns  saprogenes). 

Bacillus  foetidus  ozaenae  (Ha- 
jek). — Short  rods,  singly,  in  pairs, 
and  in  short  chains.  Motile. 

Colonies  irregular  and  liquefying. 

In  the  depth  of  gelatine  lique- 
faction occurs  along  the  track  of 
the  needle. 

On  agar  the  growth  is  moist  and 
shiny,  and  on  potato  yellowish- 
brown. 

Cultures  emit  a  disagreeable 
odour. 

They  produce  a  fatal  result  in 
mice  and  local  inflammation  in 
rabbits. 

They  were  isolated  from  cases 
of  ozasna. 

Bacillus  fulvus  (Zimmermann). 
—Rods  -88  to  1-3  fi  in  length, 
"77  fji  in  width.  Singly,  in  pairs 
and  in  chains. 

Colonies  vary  in  form  ;  granular, 
yellowish-grey. 

In  the  depth  of  gelatine  there  is 
a  scanty  faintly  yellow  growth  in 
the  track  of  the  needle,  and  a 
hemispherical  yellow  mass  on  the 
free  surface. 

On  agar  and  potato  the  growth 
is  yellow  and  shining. 

They  occur  in  water. 

Bacillus  fusCUS  (Zimmermann). 
— Rods  "63  p  in  width,  varying  in 
length  ;  sometimes  bent  and  irregu- 
lar in  form. 

Colonies  irregular,  granular,  and 
greyish-yellow . 

In  the  depth  of  gelatine  a 
hemispherical  mass  appears  on  the 
free  surface,  which  later  spreads 
and  forms  a  yellow  wrinkled  layer. 

On  agar  and  potato  the  layer  is 
similarly  coloured. 


They  occur  in  water. 
Bacillus      fuscus      limbatus 
(Scheibenzuber). — Rods    and    fila- 
ments. 

Colonies  with  brown  nucleus  and 
light  periphery. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  in  the  track  of  the 
needle  is  branching  and  the  jelly 
coloured  brown. 

On  agar  and  potato  the  growth 
is  dark  brown. 

They  occur  in  rotten  eggs. 

Bacillus  gallinarum  (Bacillus 
of  Fowl  Enteritis)  (see  p.  230). 

Bacillus  gasoformans  (Eisen- 
berg). — Rods. 

The  colonies  are  granular,  and 
liquefy  the  gelatine. 

The    bacilli    inoculated    in    the 

depth  of  gelatine  rapidly  produce 

liquefaction  in   the    track   of    the 

i  needle,     and     formation     of     gas 

bubbles. 

They  occur  in  water. 

Bacillus  gingiyae  pyogenes : 
vide  Bacterium  gingivce  pyogenes. 

Bacillus  glaucus  (Maschek).— 
Rods. 

The    colonies    are    well-defined, 
i  greyish  in  colour,  and  after  a  time 
liquefy  the  gelatine. 

The     bacilli    inoculated    in    the 

depth  of  gelatine  produce  a  rapid 

growth  in  the  track  of  the  needle 

and  on  the   surface,   followed  by 

i   liquefaction  and  a  sediment  at  the 

i   bottom  of  the  liquid. 

On  potato  and  agar  they  form  a 
1   greyish  layer. 

They  occur  in  water. 

Bacillus  gliscr  ogenus  (Malerba). 
—Rods  -57  to  1-14  p  in  length,  -41  /z 
in  width. 

Colonies  spherical,  granular. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  give  rise  to  a 
growth  in  the  track  of  the  needle 
composed  of  closely  packed  disc- 
shaped  colonies. 

On  agar  they  produce  an  opales- 
cent film. 

On  potato  they  form  a  viscid 
yellowish  growth. 

They  were  isolated  from  viscid 
urine. 

Bacillus      gracilis      (Zimmer- 

33 


514 


DESCRIPTION   OF   SPECIES. 


mann). — Rods  sometimes  curved, 
2-4  to  3-6/1  in  length,  '77  /n  in  width, 
and  filaments. 

The  colonies  are  greyish  or 
yellowish-grey,  with  concentric 
markings. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  isolated 
colonies  in  the  track  of  the  needle, 
and  a  translucent  film  on  the  free 
surface  ;  followed  after  some  time 
by  slight  liquefaction. 

On  agar  they  produce  a  bluish- 
white  layer. 

There  is  scarcely  any  growth  on 
potato. 

They  occur  in  water. 

Bacillus  gracilis  anaerobies- 
cens  (Vaughan). — Rods. 

The  colonies  are  brownish. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  copious 
growth  in  the  track  of  the  needle, 
and  gas  bubbles,  and  a  film  on  the 
surface. 

On  agar  they  produce  a  thin 
layer,  and  on  potato  a  yellowish- 
white  mass. 

They  occur  in  water. 

Bacillus  granulosus  (Russell). 
— Rods  singly,  in  pairs  and  in 
masses,  and  long  filaments.  Spore- 
formation  present. 

The  colonies  have  concentric 
linear  markings. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  cause  slow  lique- 
faction spreading  downwards  in 
the  track  of  the  needle. 

On  the  surface  of  agar  they  form 
more  or  less  isolated  whitish  or 
yellowish  colonies. 

In  broth  they  produce  turbidity, 
and  on  potato  a  thick  shining  layer, 
which  is  at  first  white  and  later 
brownish. 

They  were  isolated  from  deep-sea 
dredgings. 

Bacillus  graveolens  (Bordoni- 
Uffreduzzi). — Very  short  rods,  '8  p. 
in  length. 

The  colonies  are  greyish-white, 
and  liquefy  the  gelatine. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  rapid 
liquefaction  in  the  track  of  the 
needle.  They  colour  the  gelatine 


greenish-yellow,  and  produce  a  foul 
odour. 

On  potato  the  culture  is  brown- 
ish. 

They  were  isolated  from  skin 
from  between  the  toes. 

Bacillus  guttatus  (Zimmer- 
mann). — Rods  1  to  1/13  /t  in  length  ; 
•93  in  width  ;  singly,  in  pairs,  and 
in  chains. 

Colonies  bluish-grey  ;  granular. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  develop  colonies 
in  the  track  of  the  needle,  and  a 
greyish  opalescent  film  on  the 
surface. 

On  agar  the  growth  is  greyish- 
white.  On  potato  it  is  yellowish 
and  slimy. 

They  occur  in  water. 

Bacillus  halophilus  (Russell).— 
Rods  1-5  to  3'5  /x  in  length,  -7  p.  in 
width  ;  singly  and  in  pairs  ;  and 
toruloid  involution  forms.  Motile. 

The  colonies  liquefy  gelatine  and 
become  frothy  from  abundant  for- 
mation of  gas. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  grow  in  the  track 
of  the  needle,  and  excavate  the 
jelly  at  the  upper  part. 

They  were  isolated  from  deep- 
sea  dredgings. 

Bacillus  Hansenii  (Rasmussen). 
—Rods  2-8  to  6  p  long,  -6  to  -8  p 
wide. 

Cultivated  on  sterilised  potato, 
they  form  in  four  days  a  chrome- 
yellow  layer  with  an  agreeable 
fruit-like  smell.  Two  or  three  days 
later  the  growth  dries,  and  changes 
to  an  orange-yellow  colour  ;  later 
it  becomes  yellowish  or  brown,  and 
at  the  same  time  spores  are  formed 
1/7  p  long,  3-1  p.  wide.  The  colour- 
ing matter  is  insoluble  in  most 
reagents. 

The  bacilli  occur  as  a  yellow  or 
whitish  skin  on  nourishing  solu- 
tions, malt  infusion,  broth,  and 
wine,  which  have  been  kept  at 
31°  to  33°  C. 

Bacillus  Havaniensis  lique- 
faciens. — Rods  -8  p  in  width,  1-2 
to  5  p  in  length,  singly,  and  in 
pairs  ;  and  filaments.  Motile. 

The  colonies  are  milky,  irregular 


DESCRIPTION   OF  SPECIES. 


515 


in  outline,  and  liquefy  the  gela- 
tine. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  cause  liquefaction 
in  the  track  of  the  needle. 

On  agar  they  form  a  pale-brown 
layer. 

They  do  not  grow  on  potato. 

They  were  isolated  from  the  skin. 

Bacillus  helvolus  (Zimmer- 
mann). — Rods  1'5  to4'5  p  in  length, 
•3  p.  in  width  ;  in  pairs,  and  in 
chains. 

Colonies  are  pale  yellow. 

The  bacilli  form  a  yellow  growth 
on  the  surface  of  gelatine,  and  pro- 
duce slow  liquefaction. 

On  agar  the  growth  is  yellow. 

They  occur  in  water. 

Bacillus  heminecrobiophilus 
(Arloing). — Rods  highly  polymor- 
phic, and  filaments  1  to  20  /*  in 
length.  Slightly  motile. 

On  the  surface  of  obliquely 
solidified  gelatine  they  form  a 
yellowish  layer. 

On  potato  the  growth  is  yellowish- 
white. 

They  produce  oedema  when  sub- 
cutaneously  inoculated  in  the 
vicinity  of  wounds. 

They  were  isolated  from  a  caseous 
lymphatic  gland  in  a  guinea-pig. 

Bacillus  hepaticus  fortuitus 
(Sternberg).  —  Rods  resembling 
Bacillus  coli  communis. 

The  colonies,  marked  with  radia- 
ting strige,  are  dark  brown  in  colour. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  very 
slight  growth  at  the  upper  part  of 
the  track  of  the  needle,  and  a  hemi- 
spherical mass  on  the  free  surface. 

On  potato  they  form  a  creamy 
white  growth. 

They  were  isolated  from  the 
liver  in  a  fatal  case  of  yellow 
fever. 

Bacillus  Hessii  (Guillebeau). — 
Rods  3  to  5  /z  in  length,  1'2  /x  in 
width,  cocci-forms  and  filaments. 

The  colonies  are  filamentous, 
and  liquefy  gelatine. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  lique- 
faction, and  the  liquid  jelly  is  made 
extremely  viscous. 


On  potato  the  growth  is  brownish. 

They  coagulate  milk. 

They  were  isolated  from  milk. 

Bacillus  hyacinth!  septicus 
(Heinz).— Rods  4  to  6  /M  in  length, 
1  p.  in  width. 

The  colonies  are  transparent  and 
bluish-white. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  fila- 
ment in  the  track  of  the  needle 
and  a  layer  on  the  surface. 

On  potato  they  produce  a  slimy, 
dirty-yellow  layer. 

They  were  isolated  from  diseased 
hyacinths. 

Bacillus  hyalinus  (Jordan).— 
Rods  3*6  to  4  p.  in  length,  1*5  /*  in 
width,  and  chains. 

The  colonies  are  surrounded  by 
radiating  filaments. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  liquefac- 
tion in  the  track  of  the  needle,  u 
scum  on  the  surface  and  a  deposit 
at  the  bottom. 

On  agar  they  produce  a  dry,  grey 
growth. 

On  potato  the  growth  is  greyish- 
white  and  tuberculated. 

They  coagulate  milk. 

In  broth  they  produce  turbidity 
arid  a  pellicle  on  the  surface  ;  and 
they  are  powerful  nitrifying 
agents. 

They  occur  in  water. 

Bacillus  hydrophilus  fuscus 
(Sanarelli). — Rods  1  to  3  /t  in 
length,  and  filaments  15  to  20  p.  in 
length. 

They  rapidly  produce  a  funnel- 
shaped  area  of  liquefaction  when 
grown  in  gelatine,  followed  by 
complete  liquefaction  and  the 
formation  of  a  white  flocculent 
deposit. 

Inoculated  in  glycerine  agar  they 
"row  rapidly  and  produce  gas 
bubbles.  On  potato  they  produce 
a  straw-coloured  layer  which  be- 
comes distinctly  yellow  and  later 
brown. 

They  are  pathogenic  in  cold- 
blooded animals  and  in  small  warm- 
blooded animals.  Guinea-pigs  suc- 
cumb in  twelve  hours  ;  the  spleen 
is  enlarged  and  the  bacilli  are  found 


516 


DESCRIFriON   OF  SPECIES. 


in  great  numbers  in  the  blood  and 
internal  organs. 

They  were  isolated  from  the 
lymph  of  diseased  frogs. 

Bacillus  ianthinus  (Bacterium 
ianthinum  Zopf,  Bacillus  riolaceui). 
— Slender  rods,  about  four  times 
their  width  in  length,  with  rounded 
ends.  They  also  form  threads,  and 
are  actively  motile.  Spore-forma- 
tion present  in  the  rods. 

The  colonies  occur  as  circum- 
scribed liquefied  areas,  in  the  centre 
of  which  is  a  collection  of  the 
coloured  growth. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  funnel- 
shaped  liquefaction,  and  a  granular- 
looking  violet  mass  subsides  to  the 
bottom. 

On  agar-agar  and  potato  a 
beautiful  violet  growth  rapidly 
develops.  The  colouring  matter  is 
soluble  in  alcohol. 

They  were  observed  on  pieces  of 
pigs'  bladder  floating  on  the  surface 
of  water  rich  in  bacteria.  They 
occurred  only  on  the  surface  of  the 
1  (ladder  exposed  to  the  air.  and  never 
on  the  part  under  water.  They  occa- 
sionally occur  in  common  tap  water. 

Bacillus  implexus  (Zimmer- 
mann).  Rods  2*5  p.  in  length, 
T15  p.  in  width.  Non  -  motile. 
Spore-formation  present. 

Colonies  white,  granular,  develop- 
ing in  three  days  into  masses  of 
interlacing  white  filaments. 

Inoculated  in  the  depth  of  gela- 
tine, a  growth  develops  in  the  track 
of  the  needle  and  fine  filaments 
penetrate  the  gelatine.  The  jelly  is 
liquefied,  and  a  pellicle  forms  on 
the  surface,  and  there  is  a  flocculent 
deposit. 

On  agar  the  growth  is  white,  and 
on  potato  yellowish-white. 

They  occur  in  water. 

Bacillus  in  acne  contagiosa  in 
horses  (Dieckerhoff  and  Grawitz). 
—Short  rods  '2  /*  in  diam. 

Inoculated  in  the  depth  of 
nutrient  gelatine  they  form  a  scanty 
growth  in  the  track  of  the  needle 
and  a  white  patch  on  the  free  sur- 
face. They  thrive  best  on  blood 
serum  and  on  agar. 


The  bacilli  inoculated  on  the 
surface  of  the  skin  of  horses,  calves 
and  other  animals  are  said  to  pro- 
duce acne  pustules.  Inoculated 
subcutaneously  in  guinea-pigs  they 
produce  a  fatal  result  in  twenty- 
four  hours. 

They  were  isolated  from  pus 
in  cases  of  acne  contagiosa  in  horses. 

Bacillus  in  cancer  (Koubasoff). 
— Rods  ;  spore-formation  present. 

Inoculated  in  the  depth  of  gela- 
tine an  irregular  filament  develops 
in  the  track  of  the  needle  and  a 
transparent  growth  with  central 
depression  on  the  surface. 

They  are  said  to  be  pathogenic 
in  small  animals,  and  to  produce 
nodules  and  ulcers  of  the  mucous 
membrane  of  the  stomach. 

They  were  isolated  from  a  case 
of  cancer  e>f  the  stomach. 

Bacillus  in  cholera  in  ducks 
(Cornil  and  Toupet),  p.  230. 

Bacillus  in  choleraic  diarrhoea 
(Bovet). — Rods  2  to  4  p  in  length, 
1  to  1*5  p  in  width,  singly  and  in 
pairs,  and  filaments. 

In  the  depth  of  gelatine  a  filament 
forms  in  the  track  of  the  needle  and 
a  greyish  transparent  layer  on  the 
surface. 

On  agar  a  greyish  film  is  formed. 

On  potato  the  growth  is  yellowish 
and  abundant. 

Intra-peritoneal  injections  in 
guinea-pigs  cause  peritonitis  and 
death. 

They  were  isolated  from  a  case 
of  choleraic  diarrhoea. 

Bacillus  in  diphtheritic  disease 

of  Calves  (Bacillus  vitulonnit 
Loftier). — Rods  about  five  or  six 
times  as  long  as  wide,  mostly  united 
in  long  threads. 

A  piece  of  membrane  from  a  diph- 
theritic disease  in  a  calf,  placed  on 
blood  serum  developed  a  white  layer 
composed  of  the  bacteria.  Succes- 
sive generations  were  not  obtainable. 

Mice  inoculated  directly  from  the 
calf  died  of  a  characteristic  illness, 
and  the  same  long  bacteria  were 
again  found  in  the  inoculated 
animals  accompanying  widespread 
infiltration,  starting  from  the  point 
I  of  inoculation.  Inoculation  of 


DESCRIPTION   OF   SPECIES. 


517 


guinea-pigs  and  rabbits  gave  doubt- 
ful results.  They  were  found  in 
the  deeper  stratum  of  pseudo- 
diphtheritic  patches  in  calves. 

Bacillus  in  disease  of  bees  (p. 
471). 

Bacillus  in  erythema  node-sum 
(Demme). — Rods  2'2  to  2*5  /x  in 
length,  '5  to  '1  p.  in  width.  They 
can  be  cultivated  at  37°C. 

Colonies  on  agar  are  white  with 
radiating  lines. 

The  bacilli  inoculated  in  the  depth 
of  agar  grow  in  the  track  of  the 
needle,  and  produce  peculiar  off- 
shoots in  the  surrounding  jelly. 

They  are  said  to  produce  an 
eruption  resembling  erythema 
nodosum  when  inoculated  subcu- 
taneously  in  guinea-pigs. 

They  were  obtained  from  the 
eruption  and  the  blood  in  cases  of 
erythema  nodosum. 

Bacillus  in  fowl  enteritis 
(Klein),  p.  230. 

Bacillus  in  gangrene  (Tricomi). 
— Rods  3  ft  in  length,  I  p,  in  width, 
singly  and  in  pairs. 

Colonies  circular,  granular,  dirty- 
yellow. 

In  the  depth  of  gelatine  they 
produce  a  filament  composed  of 
closely  aggregated  colonies,  and  at 
the  upper  part  conical  liquefaction 
of  the  jelly,  beneath  a  cup- shaped 
excavation. 

On  agar  and  potato  the  growth 
is  white. 

In  jectedsubcutaneously  in  rabbits 
and  guinea-pigs  they  produce  gan- 
grene and  death  in  a  few  days. 

They  were  isolated  from  a  case 
of  senile  gangrene. 

Bacillus  in  grouse  disease 
(Klein),  p.  230. 

Bacillus  in  hog  cholera  (p. 
351). 

Bacillus  in  infantile  diarrhoea 
(Booker). —  Rods  morphologically 
identical  with  Bacillus  coli  com- 
munis.  There  are  seven  varieties 
of  this  bacillus.  They  were  iso- 
lated from  cases  of  infantile  diar- 
rhoea. 

Bacillus  in  infantile  diarrhoea 
(Lesage). — Rods  2 '4  p.  in  length, 
'75  ft  in  width,  and  filaments. 


Colonies  irregular  ia  contour, 
colouring  the  gelatine  green. 

On  the  surface  of  agar  they  form 
a  greenish  growth,  and  the  gelatine 
is  coloured  green. 

Injected  intravenously  in  a  rabbit 
they  produced  diarrhoea. 

They  are  said  to  be  identical  with 
Bacillus  fluorescens  liquefaciens 

Bacillus  in  intestinal  diph- 
theria in  rabbits  (Ribbert).— 
Rods  3  to  4  ft  in  length,  1  to  14  ft 
in  width  ;  singly,  in  pairs,  and  in 
filaments. 

Colonies  greyish  ;  granular. 

They  produce  in  gelatine  a  deli- 
cate growth  in  the  track  of  the 
needle.  They  are  pathogenic. 

They  were  isolated  from  the 
intestine  of  rabbits  suffering  from 
a  diphtheritic  inflammation  of  the 
mucous  membrane. 

Bacillus  in  jequirity  infusion 
(see  Bacillus  ofSatOer). 

Bacillus  in'measles  (p.  283). 

Bacillus  in  noma  (Schimmel- 
busch). — Rods  singly,  in  pairs,  and 
filaments. 

Colonies  circular,  greyish-white, 
granular,  with  irregular  margins. 

In   the   depth  of   gelatine   they 
produce  a  granular  filament  anda 
patch  on  the  surface. 

On  agar  and  potato  the  growth 
is  greyish-white. 

They  are  pyogenic  in  rabbits. 

They  were  inoculated  from  a 
case  of  noma. 

Bacillus  in  ophthalmia  (p.  190). 

Bacillus  in  potato  rot.— Rods 
2-5  to  4  /A  in  length,  '7  to  *8  ft  in 
width  ;  singly,  in  chains,  and  in  fila- 
ments. Spore-formation  present. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  funnel- 
shaped  area  of  liquefaction. 

On  agar  the  growth  is  composed 
of  greyish-white  slimy  colonies. 

They  were  isolated  from  diseased 
potatoes. 

Bacillus  in  purpura  haemor- 
rhagica  (Tizzoni  ana  Giovannini). 
—Rods  -75  to  1-3  ^  in  length,  "2  to 
•4  ft  in  width,  singly,  in  pairs,  and 
in  masses. 

Colonies  have  a  greyish-yellow 
nucleus  and  a  marginal  zone  of  fine 


518 


DESCRIPTION   OF   SPECIES. 


filaments  both  in  gelatine  and  agar. 
Cultures  produce  a  disagreeable 
odour. 

Subcutaneous  in  j  ections  in  guinea- 
pigs  and  rabbits  produce  local  oedema 
and  death,  with  hsemorrhages  in  the 
internal  organs. 

They  were  isolated  from  the  blood 
in  fatal  cases  of  purpura  in  children. 

Bacillus  in  putrid  bronchitis 
(Lumnitzer). — Rods  1-5  to  2  p.  in 
length,  slightly  curved.  They  can 
be  cultivated  at  37°  C.  The  colonies 
on  agar  are  greyish- white. 

The  bacilli  inoculated  on  blood 
serum  produce  colonies  which  co- 
alesce and  form  a  greyish-white 
film.  Cultures  have  a  disagreeable 
odour. 

Injected  into  the  lungs  of  rabbits 
they  produce  purulent  inflammation. 

They  were  isolated  from  the 
sputum  in  cases  of  putrid  bronchitis. 

Bacillus  in  "red-cod"  (Dautec). 
— Rods  similar  to  Bacillus  tetani, 
with  terminal  spore-formation. 

Colonies  are  circular  ;   reddish. 

On  the  surface  of  obliquely 
solidified  gelatine  they  form  a  red 
growth  in  the  track  of  the  needle 
slowly  followed  by  liquefaction. 

Cultivated  on  dried  cod  they 
produce  a  red  colour. 

They  were  isolated  from  red-cod. 

Bacillus  in  rhinoscleroma  (p. 
411). 

Bacillus  in  saliva  (Fiocca).— 
Very  short  rods  -2  to  -33  /x  in  width. 

Colonies  circular,  granular,  and 
yellowish. 

On  the  surface  of  obliquely 
solidified  gelatine  they  form  a 
growth  composed  of  transparent 
droplets. 

On  potato  they  form  a  transparent 
film. 

In  broth  flocculi  appear. 

They  are  pathogenic  in  rabbits 
and  other  small  animals,  and  they 
are  probably  a  variety  of  the  bacillus 
of  haemorrhagic  septicaemia. 

They  were  isolated  from  saliva  of 
cats  and  dogs. 

Bacillus  in  whooping  cough 
(Afanassiew). — Rods  -(j  to  2-2  p  in 
length,  singly,  in  pairs,  and  short 
chains. 


Colonies  granular,  brownish. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  scanty  growth  in  the 
track  of  the  needle  and  a  greyish 
growth  on  the  free  surface. 

On  agar  the  growth  is  greyish. 

On  potato  the  growth  is  shining 
and  yellowish  or  brownish. 

They  are  said  to  produce  symp- 
toms in  rabbits  and  dogs  compar- 
able to  those  of  whooping  cough. 

They  were  isolated  from  the 
throat  in  cases  of  whooping  cough. 

Bacillus  incanus  (Pohl).— Rods 
1*2  IJL  in  length,  "8  p,  in  width. 

They  produce  rapid  liquefaction 
in  the  track  of  the  needle  when 
inoculated  in  the  depth  of  gelatine. 

On  agar  a  thick  white  growth 
develops. 

On  potato  a  whitish  growth 
spreads  over  the  surface. 

They  were  isolated  from  the 
water  of  marshes. 

Bacillus  indicus  (Koch).— Very 
short  rods  with  rounded  ends. 


FIG.  204.— BACILLUS  INDICUS  COLONIES 
IN  NUTRIENT  AGAR,    x  60. 

The  colonies  have  a  scarlet  tint. 
They  are  round,  ovoid,  or  spindle 
shaped,  and  have  granular  margins. 

In  the  track  of  the  needle  beneath 
the  surface  no  pigment  is  formed. 

Cultivated  in  nutrient  gelatine 
they  liquefy  it  and  colour  it  crimson, 
and  the  growth  of  a  darker  crimson 
hue  subsides  to  the  bottom  of  the 
tube. 

On  the  surface  of  nutrient  agar- 
agar  the  appearances  are  very 
characteristic.  In  a  pure  cultiva- 
tion a  brilliant  vermilion-coloured 
reticulated  pellicle  develops  on  the 
surface.  (Plate  II.  Fig.  3.) 


DESCRIPTION   OF   SPECIES. 


519 


They  form  a  vermilion  layer  on 
potato. 

They  were  isolated  by  Koch  in 
India  from  the  intestinal  contents 
of  an  ape. 

Bacillus  indigogenes  (Alvarez). 
Rods  3  p  in  length  and  1'5  /u  in 
width,  singly,  and  in  chains  ;  capsu- 
lated. 

On  agar  they  produce  a  yellowish- 
white  layer,  and  are  said  to  develop 
an  indigo-blue  colour  in  infusions 
of  leaves  of  the  indigo  plant. 

Intravenous  injections  in  guinea- 
pigs  are  said  to  produce  death  in  a 
few  hours. 

They  were  isolated  from  the 
leaves  of  the  indigo  plant. 

Bacillus  indigonaceus  (Clas- 
sen).—Rods  1-6  to  3  p.  long,  -8  to 
•9  p.  wide  ;  non-motile. 

They  form  a  sky-blue  layer  on 
the  surface  of  gelatine. 

On  potato  the  growth  is  dark- 
blue,  and  later  has  a  metallic  lustre. 

Bacillus  indigoferus,  which  was 
found  in  water  at  Kiel,  is  only 
to  be  distinguished  by  its  motility. 

Bacillus  inflatus  (A.  Koch).— 
Rods  4'6  to  5'5  /A  in  length,  '6  to 
1-8  fi  in  width,  and  filaments. 

The  colonies  send  out  delicate 
processes. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  send  out  fine 
filaments  in  the  track  of  the  needle 
followed  by  slow  liquefaction. 

On  agar  they  form  a  shining 
brownish  layer. 

In  broth  a  pellicle  forms  on  the 
surface. 

They  occur  in  the  air. 

Bacillus  inunctus  (Pohl).— 
Rods  3'5  n  in  length,  -8  to  *9  /x  in 
width. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  both  in  the  track  of 
the  needle  and  on  the  surface  ; 
liquefaction  follows  in  time. 

On  agar  they  form  a  whitish 
growth. 

They  were  isolated  from  the 
water  of  marshes. 

Bacillus  invisibilis  (Vaughan). 
—Rods  ;  motile. 

The  colonies  are  irregular  and 
yellowish. 


Inoculated  in  the  depth  of  gela- 
tine they  grow  both  in  the  track  of 
the  needle  and  on  the  surface. 

On  agar  they  form  a  white 
growth. 

On  potato  they  develop  an  in- 
visible layer. 

They  occur  in  water. 

Bacillus  iridescens  (Tataroff).— 
Rods  from  3'5  to  5'2  p  in  length 
and  threads.  Spore-formation  pre- 
sent ;  slightly  motile. 

The  colonies  have  a  characteristic 
appearance  recalling  that  of  the 
convolutions  of  the  brain. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  depression  at  the 
point  of  puncture,  and  a  thread- 
like growth  along  the  needle 
track. 

On  agar  they  form  a  thick,  un- 
even, moist,  greenish-yellow,  irides- 
cent growth,  with  a  pitted  surface. 

Blood  serum  is  liquefied. 

On  potato  there  is  a  dry,  thick, 
dark  yellow  growth  like  honey. 

Broth  is  rendered  turbid,  and 
there  is  a  yellow  deposit. 

They  are  found  in  water. 

Bacillus  lactis  aerogenes  (Es- 
cherich). — Rods  short  and  thick, 
•5  to  -8  p  broad,  and  1  to  2  p  long, 
with  rounded  ends  ;  usually  in  pairs 
side  by  side  and  also  in  irregular 
heaps.  Non-motile  :  spore-forma- 
tion not  observed.  They  grow  best 
at  37°C. 

Colonies  on  the  surface  are  raised, 
moist,  shining  and  porcelain-white. 
Below  the  surface  they  have  a 
yellowish  nucleus. 

Inoculated  in  the  depth  of  gela- 
tine the  rods  form  an  abundant 
nail-shaped  growth.  On  potato  the 
culture  is  composed  of  white  colo- 
nies, and  bubbles  are  formed.  The 
colonies  may  coalesce  and  produce 
a  creamy  layer. 

On  blood  serum  there  is  a  raised, 
moist,  shining,  white  growth. 

In  milk  sugar  or  grape  sugar 
solutions  they  produce  gas. 

Injected  subcutaneously  in  rab- 
bits and  guinea-pigs  they  cause 
death  in  from  one  to  three  days, 
and  the  bacilli  are  found  in  the 
blood  and  internal  organs. 


520 


DESCRIPTION   OF   SPECIES. 


They  were  found  in  the  intestinal 
tract  of  animals  fed  with  milk  and 
of  infants  at  the  breast. 

Bacillus  lactis  albus  (Loffler). 
— Rods,  3-4  p.  in  length,  '96  /*  in 
width,  and  filaments.  Spore-forma- 
tion present. 

Inoculated  in  the  depth  of  gela- 
tine they  slowly  liquefy  the  upper 
part,  and  a  white  scum  forms  on  the 
surface. 

On  agar  they  form  a  white  layer. 

On  potato  the  growth  is  dry  and 
white.  They  coagulate  milk. 

They  occur  in  milk. 

Bacillus  lactis  erythrogenes 
(Hueppe). — Short  rods,  1  to  1/4  p, 
in  length  and  *3  to  -5  p  in  width, 
and  filaments.  Colonies  small  and 
circular  ;  greyish- white  ;  later  yel- 
low and  surrounded  by  liquefied 
gelatine  with  a  pink  tinge. 

In  the  depth  of  gelatine  the 
growth  in  the  track  of  the  needle 
is  scanty,  but  on  the  surface  a 
whitish  patch  forms  which  after- 
wards turns  yellow,  and  the  gela- 
tine is  coloured  pink.  Later  lique- 
faction sets  in,  and  the  liquefied 
gelatine  is  turbid  and  pink. 

On  agar  a  shining  yellow  layer 
develops,  and  the  same  on  potato. 

In  broth  the  bacilli  produce  tur- 
bidity, and  they  coagulate  milk. 

They  occur  in  "  red  milk." 

Bacillus  lactis  pituitosi  (Ldf- 
er). — Rods  slightly  bent. 

Colonies  circular,  greyish-white. 

On  agar  and  potato  they  pro- 
duce a  greyish- white  layer. 

They  render  milk  viscid. 

They  occur  in  milk. 

Bacillus  latericeus  (Adametz 
and  Eisenberg). — Rods  and  fila- 
ments. 

Colonies  circular,  granular,  red- 
dish-brown. 

In  the  depth  of  gelatine  there  is 
a  scanty  growth  along  the  track 
of  the  needle  and  a  brick- red 
growth  on  the  surface. 

On  potato  the  growth  is  also 
brick-red. 

They  occur  in  water. 

Bacillus  leporis  lethalis  (Gibier 
and  Sternberg). — Rods  1  to  3  p.  in 
length,  -5  /u.  in  width. 


Colonies  transparent  and  with 
the  appearance  of  broken  glass. 

In  the  depth  of  gelatine  there 
is  a  growth  along  the  track  of  the 
needle  with  a  conical  area  of  lique- 
faction at  the  upper  part,  and  a 
white  sediment. 

On  agar  they  form  a  trans- 
lucent film.  They  liquefy  blood 
serum. 

On  potato  the  growth  is  pale- 
yellow. 

Cultures  injected  into  the  peri- 
toneal cavity  of  rabbits  are  toxic. 

They  were  isolated  from  the  in- 
testinal contents  in  cases  of  yellow 
fever. 

Bacillus  leprae  (p.  407). 

Bacillus  leptosporus  (L.  Klein). 
— Rods  resembling  hay-bacilli, 
singly,  in  chains  and  long  twisted 
filaments. 

The  spore-membrane  is  said  to 
form  part  of  the  newly  grown 
bacillus,  and  the  filaments  are  de- 
scribed as  possessing  peculiar  spas- 
modic movements. 

They  were  isolated  from  a  con- 
taminated culture. 

Bacillus  limbatus  acidi  lactici 
(Marpmann).— Rods  short,  thick  : 
singly,  in  pairs  ;  capsulated. 

Colonies  white. 

In  the  depth  of  gelatine  they 
develop  slightly  in  the  track  of  the 
needle,  and  produce  a  white  patch 
on  the  free  surface. 

In  milk  they  produce  coagula- 
tion and  lactic  acid. 

They  occur  in  milk. 

Bacillus  limosus  (Russell).— 
Rods  3  to  4  p,  in  length,  1/25  \i  in 
width  ;  singly,  in  pairs  and  chains  ; 
spore-formation  present. 

Colonies  transparent,  surrounded 
by  filamentous  processes. 

In  the  depth  of  gelatine  pre- 
pared with  sea-water,  liquefaction 
occurs  rapidly  in  the  track  of  the 
needle,  and  a  deposit  forms  at 
the  bottom  and  a  thin  skin  on  the 
surface. 

On  agar  they  form  a  white  layer, 
and  in  broth  turbidity  and  a  thick 
scum. 

On  potato  the  growth  is  greyish- 
white. 


DESCRIPTION   OF   SPECIES. 


521 


They  were  isolated  from  deep- 
sea  dredgings. 

Bacillus  liodermos  (Fliigge).— 
Small  short  rods  with  rounded 
ends  ;  actively  motile. 

Colonies  with  irregular  outlines 
float  on  liquefied  gelatine  in  the 
form  of  small  white  flakes. 

Inoculated  in  gelatine  a  greyish 
growth  occurs  along  the  track  of  the 
needle,  but  the  medium  later  be- 
comes liquefied  and  a  greyish-white 
flocculent  deposit  settles  at  the 
bottom. 

On  potato  a  smooth  shining  yel- 
lowish-white layer  spreads  quickly 
over  the  whole  surface,  and  after 
some  days  becomes  opaque  and 
slightly  wrinkled. 

They  occur  on  potato. 

Bacillus  liquefaciens  (Eisen- 
berg). — Rods  short  and  thick,  with 
rounded  ends.  Very  motile. 

Colonies  round,  with  smooth 
edges  and  slimy  centres.  Lique- 
faction follows,  and  a  putrefactive 
odour  is  noticed. 

In  gelatine  they  make  a  funnel- 
shaped  whitish  growth  along  the 
track  of  the  needle. 

On  potato  the  growth  is  pale 
yellow. 

They  occur  in  water. 

Bacillus  liquefaciens  commu- 
nis  (Sternberg).— Rods  1  to  2  p  in 
length,  and  •?  //.  in  width  ;  singly 
and  in  pairs. 

In  the  depth  of  gelatine  they 
produce  rapid  liquefaction  in  the 
track  of  the  needle. 

On  potato  a  wrinkled  pinkish 
layer  is  formed. 

They  were  isolated  from  the  eva- 
cuations of  yellow-fever  patients. 

Bacillus  liquefaciens  magnus 
(Liideritz). — Rods  3  to  0  p  in 
length,  -8  to  1-1  p.  in  width,  and 
filaments.  They  are  anaerobic. 

Colonies  develop  below  the  sur- 
face of  the  gelatine,  and  liquefaction 
extends  upwards  to  the  surface. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  cause  liquefaction 
in  the  lower  part  of  the  track  of 
the  needle. 

In  the  depth  of  agar  the  colonies 
have  delicate  branches. 


They  liquefy  blood-serum,  and 
produce  a  putrefactive  odour. 

They  occur  in  earth. 

Bacillus  liquefaciens  parvus 
(Liideritz).  — -  Rods  2  to  5  n  in 
length,  -5  to  •?  M  in  width,  and 
filaments.  They  are  anaerobic. 

Colonies  are  white,  and  liquefy 
gelatine  ;  but  in  agar  they  are 
spherical  or  almond-shaped. 

In  the  depth  of  gelatine  isolated 
colonies  appear  in  the  track  of  the 
needle,  and  in  the  depth  of  agar 
there  is  gas  formation. 

They  occur  in  earth. 

Bacillus  liquidus  (Frankland). 
— Rods  short  and  flat  with  rounded 
ends,  usually  in  pairs,  the  length 
of  each  pair  varying  from  T5  pi  to 
3*5  p.  They  are  very  variable  in 
size  ;  highly  motile  ;  spore-forma- 
tion not  observed. 

Colonies  form  cup-shaped  excava- 
tions, with  almost  clear,  colourless 
contents.  The  edges  are  at  first 
smooth  and  circular,  but  they 
become  serrated  and  granular,  and 
soon  coalesce. 

A  broad  funnel-shaped  depression 
forms  along  the  whole  track  of  the 
needle,  containing  turbid  liquid  and 
masses  of  flocculent  material.  Later 
a  thin  pellicle  forms  on  the  surface, 
which  sinks  if  the  tube  is  shaken. 

On  agar  they  grow  quickly,  form- 
ing a  smooth  shining  layer. 

On  potato  a  thick  flesh-coloured 
growth  appears. 

Broth  is  rendered  turbid  with  an 
abundant  sediment,  and  after  a  few 
days  a  pellicle  forms. 

They  are  common  in  unfiltered 
water. 

Bacillus  litoralis  (Russell).— 
Colonies  granular,  with  regular 
contour  ;  slowly  liquefying. 

In  the  depth  of  gelatine  they 
develop  a  growth  in  the  track  of 
the  needle,  and  at  the  upper  part 
produce  a  cup-shaped  cavity  lined 
with  the  culture.  The  gelatine  is 
tinged  with  brown  in  the  vicinity. 

On  agar  they  produce  a  greyish- 
white  film,  and  in  broth  turbidity. 

Inoculated  in  the  depth  of 
gelatine  the  bacilli  form  a 
funnel-shaped  liquefaction  along 


522 


DESCRIPTION   OF   SPECIES. 


the  track  of  the  needle,  and  the 
whole  of  the  gelatine  gradually 
becomes  liquid,  with  a  flocculent 
deposit  at  the  bottom  and  a  greyish 
wrinkled  stain  on  the  surface. 

On  potato  a  thick  wrinkled  whitish 
skin  forms,  which  rapidly  grows 
over  the  whole  surface.  On  at- 
tempting to  raise  this  skin  it  will 
be  found  to  be  attached  to  the 
potato  by  a  mucous  substance 
which  may  be  drawn  out  in  long 
threads.  According  to  Hueppe  the 
bacilli  cannot  form  any  ropy  sub- 
stances from  sugar,  but  they  have 
an  energetic  diastatic  action.  They 
coagulate  the  casein  in  milk  in  a 
similar  manner  to  rennet. 

The  bacilli  are  ubiquitous. 

Bacillus  lividus  (Plagge  and 
Proskauer) .  — Rod  s . 

Colonies  blue-black,  liquefying. 

In  the  depth  of  gelatine  they 
produce  a  colourless  thread  in  the 
track  of  the  needle  and  a  violet 
layer  on  the  surface  followed  by 
gradual  liquefaction. 

On  agar  the  growth  is  blue-black, 
and  on  potato  violet. 

They  were  isolated  from  water. 

They  are  probably  identical  with 
Bacillus  ianthinus,  or  merely  a 
variety. 

Bacillus  luteus  (Fliigge).  — 
Short  immotile  rods. 

Colonies  irregular  in  form,  appear 
brownish  under  a  low  power,  and 
yellow  to  the  naked  eye. 

In  test-tube  cultivations  they 
form  a  yellow  growth  without 
liquefying  the  gelatine. 

They  occur  contaminating  plate- 
cultivations. 

Bacillus  maidis  (Cuboni).  - 
Rods  2  to  3  p.  in  length,  singly, 
in  pairs,  and  in  chains ;  spore- 
formation  present. 

Colonies  granular,  with  wrinkled 
periphery  ;  later,  liquefying. 

In  the  depth  of  gelatine  they 
produce  rapid  liquefaction  in  the 
track  of  the  needle. 

On  agar  a  dry  wrinkled  white 
film  spreads  over  the  surface. 

On  potato  the  growth  is  wrinkled, 
and  later  yellowish-brown.  They 
liquefy  blood  serum. 


They  were  isolated  from  human 
evacuations  and  infusions  of  maize. 

Bacillus  mallei  (p.  452). 

Bacillus  megatherium  (De 
Bary). — Rods  2 -5  p.  wide  and  four 
to  six  times  as  long,  with  rounded 
ends  and  slightly  curved,  and  in 
short  irregular  chains.  Transverse 
division  .  occurs,  each  segment 
attaining  the  length  of  the  original 
rod.  In  the  fresh  state  they  appear 
non-articulated,  but  when  treated 


FIG.  205.— BACILLUS  MEGATHERIUM. 
(a)  A  chain  of  rods  x  250,  the  rest 
x  600.  (b)  Two  active  rods :  d  and  /, 
successive  stages  of  germination ;  h 
and  I,  successive  stages  of  germina- 
tion. (De  Bary. ) 

with  a  dehydrating  agent  they  are 
seen  to  be  composed  of  short  seg- 
ments with  granular  contents.  They 
are  motile. 

Colonies  are  small  and  circular, 
and  the  gelatine  is  liquefied. 

In    the    depth    of    gelatine   the 
bacilli    grow    rapidly,    forming    a 
|   funnel-shaped  liquefaction  in   the 
!   upper  part. 

On    agar    they   form   a   whitish 
layer  on  the  surface,  and  the  jelly 
'   acquires  a  dark  colour. 

On  potato  yellowish-white  cheesy 

colonies  are  formed  round  the  point 

I   of  inoculation.     In   cultures  there 

I   is  copious   spore-formation.     They 

;   grow  best  at  20°  C. 

They    were     isolated    originally 
from  boiled  cabbage. 


DESCRIPTION    OF   SPECIES, 


523 


FIG.  206. — PURE-CULTURE  OF  BACILLUS 
MEGATHERIUM  IN  GELATINE. 


Bacillus  membranaceus  ame- 
thystinus  (Eisenberg). — Short  rods 
with  rounded  ends  from  1  to  1*4  fj. 
long,  and  "5  to  '8  \i  broad.  They 
are  grouped  together  irregularly. 
Some  individual  bacilli  stain  more 
deeply  at  the  ends  than  in  the 
middle.  Non-motile.  They  grow 
only  between  15°  and  20°  C.  Spore- 
formation  uncertain. 

The  colonies  gradually  assume  a 
violet  hue,  and  after  liquefying  the 
gelatine  float  on  the  surface  as  violet 
pellicles,  resembling  a  membrane 
stained  with  gentian  violet. 

Inoculated  in  the  depth  of 
gelatine  a  yellowish -white  growth 
appears  on  the  free  surface,  which 
after  ten  days  or  more  becomes 
violet.  Liquefaction  takes  place 
gradually,  and  in  about  a  month 
a  thick  violet  layer  covers  the  gela- 
tine which  lies  beneath  the  liquid 
part. 

On  agar  the  growth,  which  at  first 
has  a  yellowish  milky  appearance, 
becomes  violet  after  eight  or  ten 
days.  In  three  or  four  weeks  it 
has  become  very  much  wrinkled, 
and  has  a  beautiful  deep-violet 
colour  with  a  metallic  lustre.  The 


jelly  is  not  stained,  and  the  growth 
can  be  easily  removed  from  its 
surface. 

On  potato  they  grow  slowly,  and 
form  a  dirty  yellow  or  olive-green 
colour. 

In  broth  they  grow  very  slowly  ; 
after  some  weeks  a  violet  deposit 
and  pellicle  are  formed,  and  the 
liquid  between  becomes  dark  brown. 

They  were  found  in  well  water. 

Bacillus  meningitidis  puru- 
lentae  (Neumann  and  Schaffer). — 
Rods  2  /z  in  length,  -6  to  "1  p.  in 
width,  and  filaments. 

Colonies  granular,  greyish. 

In  the  depth  of  gelatine  a  greyish- 
yellow  filament  develops,  composed 
of  closely  packed  colonies,  and  on 
the  surface  a  greyish  layer. 

On  potato  the  growth  is  moist 
and  white. 

They  are  pyogenic  in  small  ani- 
mals and  dogs. 

They  were  isolated  from  a  case 
of  purulent  meningitis. 

Bacillus  mesentericus  fuscus 
(Fliigge). — Rods  small  and  short, 
singly,  in  chains  of  two  and  four. 
Actively  motile.  Spore-formation 
present. 

Colonies  are  at  first  roundish  and 
rather  white,  with  a  sharp  outline  : 
later  delicate  brownish-yellow  pro- 
cesses appear.  Liquefaction  occurs 
rapidly- 
Inoculated  in  the  depth  of  gela- 
tine a  whitish  growth  forms  along 
the  track  of  the  needle,  the  upper 
portion  of  which  soon  liquefies ; 
greyish  flakes  float  in  the  liquefied 
portion. 

On  potato  a  smooth  yellowish 
growth  appears  on  the  first  day, 
but  it  soon  becomes  brown  and 
wrinkled.  It  remains  relatively 
thin  and  superficial,  and  quickly 
spreads  over  the  whole  surface. 

They  are  found  in  hay  dust,  in 
the  air,  on  the  surface  of  potatoes, 
and  are  very  widely  distributed. 

Bacillus  mesentericus  ruber.— 
Slender  rods,  singly,  in  pairs,  and 
in  filaments. 

Colonies  are  circular  and  yellow- 
ish until  they  come  to  the  surface, 
when  they  produce  a  network  and 


524 


DESCRIPTION   OF   SPECIES. 


liquefy  the  gelatine.  The  network 
disappears  and  a  little  deposit  occurs 
at  the  bottom  of  the  liquefied 
area. 

Inoculated  in  the  depth  of  gela- 
tine a  whitish  cloudy  growth  forms 
along  the  needle  track,  liquefaction 
sets  in  and  extends  until  the  gela- 
tine is  completely  liquefied. 

On  potato  a  thin  crinkled  film  is 
formed,  which  is  yellowish  or  red- 
dish-yellow in  colour. 

They  occur  on  potato. 

Bacillus  mesentericus  vulga- 
tllS  (Fliigge). — Rods  large  and 
thick,  of  ten  forming  pseudo-threads. 
They  have  an  oscillating  movement. 
Spore-formation  present. 

The  colonies  are  bluish-white  and 
almost  transparent,  though  the 
centres  become  gradually  opaque. 
They  sink  in  the  liquefied  gelatine, 
and  are  granular  with  irregular 
contour. 

Bacillus  multipediculus 
(Fliigge). — Rods  long  and  slender. 
Non-motile. 

The  colonies  consist  of  a  central 
oval  nucleus,  from  which  numerous 
tapering  processes  shoot  out  mostly 
towards  one  pole.  This  form  of 
growth  gives  a  curious  resemblance 
to  an  insect  with  feet  and  antennse. 

Inoculated  in  the  depth  of  gela- 
tine a  whitish  line  forms  along  the 
track  of  the  .needle,  from  which 
short  processes  grow  out. 

On  potato  a  rather  scanty  dirty- 
yellow  growth  forms,  and  the  sur- 
face of  the  potato  becomes  dis- 
coloured around  it. 

They  are  often  found  as  a  con- 
tamination on  potato. 

Bacillus  muscoides  (Liborius) 
— Rods  1  p,  thick,  sometimes  form- 
ing threads  ;  slightly  motile,  and 
with  round  or  oval  spores  at  one 
end.  They  are  anaerobic. 

The  colonies  ramify  and  resemble 
a  delicate  moss. 

They  were  found  in  the  oedema- 
tous  fluid  of  field  mice  inoculated 
with  garden  earth  and  stale  cheese. 
Bacillus  mycoides  (Fliigge).— 
Rods  rather  thick,  nearly  the  size 
of  Bacillus  arithracis.  Motile, 
often  forming  long  pseudo-threads. 


Oval  and  highly  refractive  spores 
both  in  the  rods  and  threads. 

Colonies  consist  of  a  whitish  tur- 
bidity in  which  colourless  branched 
and  interwoven  processes  are  seen  ; 
these  increase  rapidly,  and  after 
twelve  to  twenty  hours  appear  like 
the  mycelium  of  a  fungus. 

Inoculated  in  the  depth  of  gela- 
tine they  form  very  fine  and  closely 
set  hairs  extending  from  the  track 
of  the  needle.  Later  liquefaction 
occurs. 

On  potato  a  whitish  layer  gradu- 
ally extends  over  the  surface. 

They  occur  in  earth  from  the 
surface  of  cultivated  ground. 

Bacillus  mycoides  roseus 
(Scholl).— Rods. 

Colonies  composed  of  interlacing 
filaments. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  in 
the  track  of  the  needle  ;  a  reddish 
scum  forms  on  the  surface,  and  a 
reddish  deposit  at  the  bottom  of 
the  liquefied  area. 

On  agar  they  produce,  in  the 
absence  of  light,  a  pink  growth. 

Bacillus  neapolitanus  (Emme- 
rich).— Short  rods  '9  fj.  in  width. 


FIG.  207. — BACILLUS  NEAPOLITANUS,  x 
700  (EMMEKICH).  a,  From  intes- 
tinal contents  in  a  case  of  cholera  ; 
b,  From  peritoneal  fluid  of  an 
inoculated  guinea-pig. 


DESCRIPTION    OF   SPECIES. 


525 


Colonies  circular,  later  irregular, 
granular,  strongly  refractive  and 
yellowish  -  brown.  They  are 

probably  identical  with  Bacillus  coli 
communis. 

They  were  isolated  from  cases  of 
cholera  at  Naples. 

Bacillus  necrophorus  (Loffler). 
— Rods  and  filaments. 

They  cannot  be  cultivated  on  the 
ordinary  media.  In  rabbit  broth 
they  give  rise  to  fluify  masses  of 
filaments. 

Intravenous  injection  produced 
in  rabbits  a  pysemic  condition  in 
about  a  week.  The  bacilli  were 
found  in  the  pus. 

They  were  isolated  from  a  rabbit 
which  had  been  inoculated  with 
fragments  of  a  condyloma. 

Bacillus  nitrificans  (Wino- 
gradsky). — Very  small  rods  -5  /*  in 
in  length,  singly  and  in  zooglcea. 

Colonies  in  silica  jelly  are 
lenticular,  and  sub-cultures  in  liquid 
media  produce  a  gelatinous  de- 
posit. They  are  powerful  oxidising 
agents. 

They  were  isolated  from  the  soil. 

Bacillus  nodosus  parvus 
(Lustgarten). — Rods  1-2  to  %2'4  p  in 
length,  -4  p  in  width  ;  singly  and 
in  pairs. 

Inoculated  in  the  depth  of  agar 
they  produce  a  white  filament  in 
the  track  of  the  needle  composed 
of  crowded  colonies,  and  on  the 
surface  a  hemispherical  glistening 
growth. 

They  were  isolated  from  the 
human  urethra. 

Bacillus  nubilus(Frankland). — 
Slender  rods  3  p.  long  and  '3  /z  wide, 
and  threads.  Single  bacilli  have  an 
active  rotatory  movement,  but  the 
long  threads  in  broth  cultures  are 
quite  motionless.  Spore-formation 
not  observed. 

The  colonies  appear  as  cloudy 
undefined  patches,  which  rapidly 
liquefy  the  gelatine.  They  consist 
of  a  tangled  mass  of  threads. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  along  the  track 
of  the  needle  horizontal  circular 
plates,  with  a  delicate  cloud-like 
appearance,  and  liquefaction  at  the 


upper  part.  Later  the  whole  of 
the  gelatine  is  liquefied. 

On  agar  they  form  a  thin  opales- 
cent blue-violet  film,  the  edges  of 
which  exhibit  later  a  distinct  violet 
fluorescence. 

On  potato  there  is  a  slightly 
yellow  growth  which  is  scarcely 
visible. 

Broth  is  rendered  turbid  with  a 
dirty-white  deposit,  the  surface 
being  covered  by  a  thin  pellicle. 

They  occur  in  water. 

Bacillus  ochraceus  (Zimmer- 
mann). — Rods  1-25  to  4*5  \i  in 
length ;  -65  to  '75  \L  in  width  ; 
singly,  in  pairs,  chains,  and  fila- 
ments ;  capsulated. 

Colonies  circular,  granular,  yel- 
low, liquefying. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  in 
the  track  of  the  needle,  and  a. 
yellow  deposit. 

On  agar  and  potato  the  growth 
is  yellow  ochre  in  colour. 

They  occur  in  water. 

Bacillus  oedematis  aerobicus 
(Klein).— Rods  '8  to  2-4  /x  in  length, 
•7  fj.  wide,  and  long  filaments. 

Colonies  greyish,  transparent, 
with  irregular  contour. 

In  the  depth  of  gelatine  a  fila- 
ment occurs  in  the  track  of  the 
needle,  and  gas  bubbles  in  isolated 
colonies  in  its  lower  part,  and  a 
transparent  patch  with  irregular 
margin  on  the  free  surface. 

On  the  surface  of  agar  they  pro- 
duce a  greyish- white  layer. 

In  broth  there  is  turbidity  with 
I  flocculi. 

On  potato  the  growth  is  yellowish 
and  viscid. 

They  give  rise  to  extensive  oedema 
in  guinea-pigv«i,  and  in  a  less  marked 
!  form  in  rabbits. 

They  occur  in  earth. 

Bacillus  oedematis  maligni 
!  (p.  2-20). 

Bacillus  of  Belfanti  and  Pas- 
car  Ola.—  Very  short  rods. 

Colonies  circular,  granular,  yel- 
i  lowish-grey. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  a  filament  com- 
;  posed  of  closely-packed  minute 


52(> 


DESCRIPTION   OF   SPECIES. 


colonies,    and    on    the    surface    a 
greyish  film. 

On  agar  they  produce  a  greyish- 
white  growth. 

On  potato  a  transparent  whitish 
film. 

They  are  fatal  to  rabbits,  guinea- 
pigs  and  small  birds. 

They  are  probably  identical  with 
Bacillus  septicaemias  haemorrhagicae. 
They  were  isolated  from  pus  in 
a  case  of  tetanus. 

They  were  isolated  from  deep-sea 
dredgings. 

Bacillus  of  Colomiatti.— Minute 
rods.  Spore-formation  occurs  at 
the  ends  of  the  rods.  They  can 
be  cultivated  at  37°  C. 

They  form  a  thin  film  on  agar 
and  on  blood  serum. 

They  were  isolated  in  cases  of 
conjunctivitis. 

Bacillus  of  Fulles,  No.  I.— Rods 
1  to  1*2  p.  in  length,  -6  /z  in  width. 

Colonies  circular,  granular,  yel- 
lowish-brown. 

On  the  surface  of  gelatine  they 
produce  a  thin  film,  and  in  broth 
turbidity  and  flocculi. 

On  potato  the  growth  is  yel- 
lowish. 

No.  II.  Very  short  rods. 
Colonies  circular,  granular,  yel- 
lowish. 

In  the  depth  of  gelatine  the 
growth  resembles  Friedlander's 
pneumococcus. 

On  potato  the  growth  is  yellowish. 
They  were  isolated  from  earth. 
Bacillus  of  Ghiillebeau.— No.  I. 
Short  rods  1  to  2  ft  in  length,  1  ^ 
in  width. 

Colonies  spherical,  granular. 
In  gelatine  the  bacilli  produce  a 
growth  in  the  track  of  the  needle 
and  a  white  patch  on  the  surface. 

On  agar  the  growth  is  white,  and 
on  potato  yellowish,  viscid,  and  con- 
taining gas  bubbles. 
They  coagulate  milk. 
No.    II.    Rods    resembling    the 
above  described  but  distinguished 
by  the  production  of  viscid  colonies 
and,  extremely  slowly,  of  liquefac- 
tion in  the  jelly. 

No.  III.  Rods  also  resembling  the 
above  mentioned,  but  colonies  are 


adherent  to  the  jelly  and  coarsely 
granular. 

Milk  and  other  liquid  culture 
media  are  rendered  extremely  vis- 
cid. 

Bacillus    of   Letzerich.— Rods 
sometimes  bent,  and  filaments. 
They  rapidly  liquefy  gelatine. 
They  produce  purulent  peritonitis 
and  death  in  rabbits. 

They  were  isolated  from  urine. 
Bacillus   of  Martinez  (Stern- 
berg). — Short   rods  1   to  1*2  ft   in 
length  and  "5    to    '8  /z  in  width  ; 
non-motile. 

Colonies  circular  and  translucent, 
with  a  central,  nipple-like  projec- 
tion, and  the  surface  covered  with 
mosaic  markings. 

In  the  depth  of  gelatine  the 
growth  consists  of  large  spherical 
translucent  colonies  in  the  track  of 
the  needle,  and  a  thin,  translucent, 
scanty  growth  upon  the  surface. 

They  were  isolated  from  the  liver 
in  a  fatal  case  of  yellow  fever. 

Bacillus  of  Nocard.  (Vide  Strep, 
tothrix  farcinica.) 

Bacillus  of  Okada.— Short  rods 
rather  thicker  than  the  bacilli  of 
mouse-septicasmia,  singly,  in  pairs 
and  in  filaments.  Spore-formation 
not  observed. 

Colonies  granular  and  brownish. 

Inoculated  in  the  depth  of  gelatine 
they  form  a  white  filament,  and  on 
the  surface  a  milk-white  patch. 

Inoculated  on  agar  the  growth 
spreads  over  the  surface  forming  a 
milk-white  layer. 

In  broth  they  produce  cloudiness 
and  a  layer  floating  on  the  surface. 

They  do  not  grow  on  potato. 

Cultures  produce  death  in  mice, 
guinea-pigs  and  rabbits  in  twenty 
hours. 

They  were  isolated  from  dust. 

Bacillus  of  Koth.— No.  1  and 
No.  2.  Rods. 

Two  varieties  were  isolated  from 
old  rags.  They  appear  to  be 
varieties  of  Bacillus  coli  communis. 

Bacillus  of  Sattler.— 2  to  4-5  /* 
long  and  '58  p  thick. 

They  can  be  cultivated  on  nutrient 
gelatine  and  blood  serum. 

Infusion  of  jequirity  containing 


DESCRIPTION   OF   SPECIES. 


527 


the  bacilli,  inoculated  into  the  con- 
junctiva of  healthy  rabbits,  produces 
severe  ophthalmia.  The  poisonous 
principle  is  a  chemical  ferment 
dbrln.  Boiling,  which  does  not 
destroy  the  spores  of  the  bacillus, 
destroys  the  ferment,  and  cultiva- 
tions started  from  these  spores, 
though  teeming  with  jequirity 
bacilli,  are  quite  harmless  (Klein). 

The  bacilli  occur  in  infusions 
of  the  beans  of  Abrus  precatorius 
or  jequirity. 

Bacillus  of  Schaffer  (Freuden- 
reich). — Rods  2  to  3  /u  in  length, 
1  fjt  in  width,  and  long  filaments. 

Colonies  circular,  granular,  yel- 
lowish. 

In  the  depth  of  gelatine  a  growth 
develops  in  the  track  of  the  needle 
and  a  greyish  layer  on  the  surface. 

On  agar  the  growth  is  greyish 
and  sometimes  brownish,  and  on 
potato  yellowish. 

In  broth  with  peptone  and  milk 
sugar  there  is  copious  formation  of 
gas-bubbles. 

They  closely  resemble  Bacillus 
coli  communis. 

They  were  isolated  from  cheese 
and  potato. 

Bacillus  of  Scheurlen.— Rods 
T5  to  2*5  p,  in  length,  '5  /*  in  width. 
They  were  isolated  from  cancerous 
growths  by  Scheurlen,  and  were 
later  identified  with  Bacillus  epider- 
midis. 

Bacillus  of  Schou. — Short  rods 
and  cocci-forms. 

Colonies  are  spherical,  opaque, 
and  granular. 

The  bacilli  inoculated  in  gelatine 
rapidly  liquefy  it,  and  a  white  de- 
posit forms  at  the  bottom  of  the 
liquid. 

Rabbits  inoculated  in  the  trachea, 
or  made  to  inhale  pure-cultures,  are 
said  to  develop  fatal  pneumonia. 
They  were  isolated  from  rabbits 
with  pneumonia,  following  section 
of  the  vagi. 

Bacillus  of  swine  plague 
(p.  351). 

Bacillus  of  Tommasoli.— Short 
rods  from  1  to  1*8  /*  in  length,  and 
•25  to  '3  /x  in  width,  singly,  and  in 
short  chains. 


Colonies  grey  and  shiny. 

In  the  depth  of  gelatine  they 
form  a  filament  composed  of  closely- 
packed  colonies,  and  on  the  surface 
a  shining  mass. 

On  agar  the  growth  consists  of 
greyish  patches. 

On  potato  the  growth  is  granular 
and  yellowish-white. 

Cultures  rubbed  into  the  skin 
are  said  to  produce  a  vesicular 
eruption. 

They  were  isolated  from  the 
scalp  in  a  case  of  sycosis. 

Bacillus  of  Utpadel.— Rods  1-25 
to  1'5  a  in  length,  and  "75  to  1  /u 
in  width,  singly,  in  pairs  and  short 
chains. 

Colonies  milk-white. 

On  the  surface  of  gelatine  the 
growth  is  milk-white,  and  on  agar 
yellowish- white . 

Injected  subcutaneously  in  cats, 
guinea-pigs  and  mice,  they  produce 
extensive  oedema,  and  a  fatal  ter- 
mination. 

They  were  isolated  from  the 
human  intestine. 

Bacillus  of  Winogradsky.  (See 
Bacillus  nitrificans.) 

Bacillus  ovatus  minutissi- 
mus  (Unna).  —  Short  rods  with 
pointed  ends  '6  to  '8  p  in  length, 
•4  ju.  in  width,  singly,  and  in 
masses. 

Colonies  are  minute,  granular  and 
yellowish. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  form  a  filament 
of  closely  packed  greyish-white 
colonies,  and  on  the  free  surface 
there  is  a  shiny,  greyish- white 
layer. 

On  agar  the  growth  is  very  simi- 
lar, and  on  potato  also. 

They  were  isolated  from  the  skin 
in  eczema  seborrhceicum. 

Bacillus  oxytocus  perniciosus 
(Wyssokowitch). — Rods  short  and 
thick. 

Colonies  circular,  granular,  yel- 
lowish, or  yellowish-brown. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  growth 
resembling  Friedlander's  pneumo- 
coccus. 

They  coagulate  milk. 


528 


DESCRIPTION    OF   SPECIES. 


The  products  injected  intrave- 
nously produce  death  in  from  three 
to  twenty-four  hours. 

They  occur  in  sour  milk. 

Bacillus  pestifer  (Frankland). 
—Rods  2-3  p.  in  length,  1  ^  in 
width,  and  filaments.  Motile. 

Colonies  resemble  those  of  Bacil- 
lus vermicularis. 

On  agar  they  produce  a  dentated 
transparent  layer,  and  on  potato  a 
flesh-coloured  growth. 

They  occur  in  the  air. 

Bacillus  phosphor escens  geli- 
dus  (Forster). — Very  short  rods. 

Colonies  circular,  granular,  yel- 
lowish or  greenish. 

The  bacilli,  inoculated  in  the  depth 
of  gelatine,  produce  very  little 
.  growth  in  the  track  of  the  needle, 
and  a  white  film  on  the  surface. 

On  agar  and  potato  the  growth 
is  whitish. 

Cultures  are  photogenic. 

They  were  isolated  from  phos- 
phorescent fish. 

Bacillus  phosphorescens  Indi- 
CUS  (Fischer). — Rods  singly  and  in 
pairs,  and  filaments.  Motile. 

Colonies  circular,  well-defined, 
greenish. 

The  bacilli,  inoculated  in  the 
depth  of  gelatine,  produce  a  greyish 
filament  in  the  track  of  the  needle, 
and  a  hemispherical  excavation  of 
the  jelly  at  the  upper  part.  Later 
the  jelly  is  liquefied,  and  there  is  a 
yellowish  scum  on  the  surface. 

On  agar  and  potato  the  growth 
is  white. 

Cultures  are  photogenic. 

They  were  isolated  from  sea- 
water. 

Bacillus  phosphorescens  indi- 
genus  (Fischer). — Rods  1-3  to  1'2 
p.  in  length,  €4  to  '7  /*  in  width, 
singly,  in  pairs,  and  filaments. 

Colonies  circular,  greenish,  and 
later  yellowish. 

The  bacilli,  inoculated  in  the 
depth  of  gelatine,  produce  a  conical 
excavation  in  the  upper  part  of  the 
needle  track  without  liquid  con- 
tents, but  with  a  dry  growth  on 
the  sides. 

There  is  no  growth  on  potato. 

Cultures  are  photogenic. 


They  occur  in  sea-water  and  on 
phosphorescent  fish. 

Bacillus  plicatus  (Zimmer- 
mann). — Minute  rods,  singly,  in 
pairs,  and  in  short  chains. 

Colonies  yellowish-white. 

The  bacilli,  inoculated  in  the 
depth  of  gelatine,  form  minute 
isolated  colonies,  and  on  the  surface 
a  wrinkled  patch  with  gradual  lique- 
faction. 

On  potato  the  growth  is  dry  and 
yellowish. 

They  occur  in  water. 

Bacillus  pneumosepticus 
(Babes). — Short  rods,  '2  p  in  width. 

Colonies  irregular,  semi  -  trans- 
parent. 

In  gelatine,  the  bacilli  grow  in 
the  track  of  the  needle.  On  agar 
the  growth  is  whitish  and  shining. 

Rabbits,  guinea-pigs  and  mice 
die  in  two  or  three  days  of  septi- 
caemia when  a  culture  is  injected 
subcutaneously. 

They  were  isolated  from  a  fatal 
case  of  septic  pneumonia. 

Bacillus  polypiformis  (Libo- 
rius). — Slender  rods,  spore-forma- 
tion present.  They  are  anaerobic. 

Colonies  composed  of  peculiar 
convoluted  processes. 

In  the  depth  of  blood  serum  they 
produce  a  cloudiness  at  the  lower 
part  of  the  needle  track. 

They  occur  in  soil. 

Bacillus  prodigiosus  (Micro- 
coccus  prodigiosus  :  Cohn. — Blood 
rain,  Bleeding  host).  Very  short 
rods  with  rounded  ends,  and  thread 
forms  *5  to  1  /u,  in  width,  forming 
at  first  rose-red  and  then  blood-red 
zoogloea. 

They  liquefy  gelatine. 

They  grow  luxuriantly  on  the 
sloping  surface  of  nutrient  agar- 
agar,  and  on  sterilised  potato,  and 
the  colour  varies  from  blood-red  to 
bright-red  with  sometimes  a  metal- 
lic lustre.  The  cells  themselves- 
are  colourless.  The  colouring-matter 
resembles  f  uchsine  ;  it  is  insoluble 
in  water  but  soluble  in  alcohol. 
The  addition  of  acids  changes  it  to 
carmine  red,  and  of  alkalies  to  a 
yellow  colour. 

They     appear     occasionally     on 


DESCRIPTION   OF   SPECIES. 


529 


bread,  boiled  rice,  and  starch  paste, 
and  more  rarely  on  boiled  white 
of  egg  and  meat.  Milk  sometimes 
becomes  coloured  blood-red  by  the 
growth  of  this  fungus,  an  appear- 
ance formerly  attributed  to  a  disease 
of  the  cow. 

In  Paris  in  1843  the  micro- 
organism was  peculiarly  prevalent, 
attacking  especially  the  bread  pro- 
duced in  the  military  bakehouses. 

Bacillus  proteus  fluorescens 
(Jager). — Short  thick  rods  and 
threads.  Actively  motile. 

Colonies  resemble  minute  drops 
of  water. 

The  rods  inoculated  in  gelatine 
produce  a  growth  similar  to  that 
of  Koch's  comma-bacilli. 

The  jelly  becomes  greenish,  and 
a  pellicle  forms  on  the  surface. 

On  agar  the  growth  when  fully 
developed  is  yellowish- white,  with 
a  green  fluorescence. 

On  potato  they  form  a  brown 
layer. 

They  are  pathogenic  in  mice. 

They  were  isolated  from  the 
internal  organs  of  fowls  suffering 
from  an  epidemic  disease. 

Bacillus  pseudo-diphtheriticus 
(p.  335). 

Bacillus  pseudo-tuberculosis 
(Pf  eiffer). — Rods  varying  in  length. 

Colonies  circular,  with  dark  nu- 
cleus and  transparent  zone. 

In  the  depth  of  gelatine  they 
produce  a  filament  composed  of 
small  colonies,  and  on  the  surface 
a  patch  with  concentric  markings. 

They  grow  on  agar,  but  not  readily 
on  potato. 

Inoculated  in  mice,  guinea-pigs, 
rabbits,  and  hares,  they  produce  a 
fatal  result  in  from  six  to  twenty 
days.  An  abscess  forms  locally, 
the  lymphatic  glands  enlarge  and 
caseate,  and  the  internal  organs 
contain  nodules  resembling  tubercle. 

They  were  isolated  from  the  in- 
ternal organs  of  a  horse  supposed 
to  be  glandered. 

Bacillus  pulpae  pyogenes.— 
Rods  slightly  bent  and  with  pointed 
ends  ;  singly,  in  pairs,  and  in  chains. 

Colonies  circular,  yellowish- 
brown. 


Inoculated  in  the  depth  of  gelatine 
liquefaction  occurs  in  the  upper 
part  of  the  needle  track  and  ex- 
tends downwards. 

Intraperitoneal  injection  in  mice 
produces  death  in  from  eighteen  to 
thirty-six  hours. 

They  were  isolated  from  putrid 
dental  pulp. 

Bacillus  punctatus  (Zimmer- 
mann). — Rods  1  to  1-6  in  length, 
•77  p.  in  width,  singly,  in  pairs,  and 
chains. 

Colonies  composed  of  stringy 
masses  in  liquefied  gelatine. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  rapid 
liquefaction  in  the  track  of  the 
needle,  and  a  white  deposit. 

On  agar  the  growth  is  smooth 
and  shining. 

On  potato  the  growth  is  brownish. 

They  occur  in  water. 

Bacillus  putrificus  coli  (Bien- 
stock). — Slender,  motile  rods,  3  //. 
in  length,  often  less,  sometimes 


FIG.  208. — BACILLUS  PUTRIFICUS  CoLir 

X    1000  (BlENSTOCK). 

forming  long  threads.  Spore-for- 
mation present. 

Cultivations  in  gelatine  are  iri- 
descent. 

They  are  constantly  present  in 
faeces. 

Bacillus  pyocyaneus  (Gessard). 
— Slender  rods,  singly,  in  twos 
and  threes,  or  in  irregular  masses. 
Spore-formation  present. 

White  colonies  appear  in  twenty- 
four  hours,  which  liquefy  the  gela- 
tine. The  whole  of  the  medium 
acquires  a  greenish  shimmer. 

If  the  bacilli  are  cultivated  in 
gelatine,  the  jelly  is  liquefied,  and 
coloured  green  by  reflected  light,  and 
a  deep  orange  by  transmitted  light. 

On  agar  they  form  a  white  layer, 
and  colour  the  medium  a  pea-green. 

34 


530 


DESCRIPTION   OF   SPECIES. 


On  potato  a  dry  rust-brown 
growth  appears  at  the  seat  of 
inoculation,  which  becomes  green 
when  treated  with  ammonia. 

The  pigment  formed  by  the  micro- 
organism is  a  definite  principle — 
pyocyanin.  It  can  be  extracted 
with  chloroform  from  pus  and  from 
washing  of  bandages  ;  it  is  soluble 
in  acidulated  water,  which  it  colours 
red.  In  neutral  solution  it  becomes 
blue.  It  crystallises  in  chloroform 
in  long  needles  ;  and  forms  some- 
times lamellae  and  prisms. 

They  cause  death  in  guinea-pigs 
wh£h  injected  into  the  abdominal 
cavity.  Eabbits  are  not  killed  by 
intravenous  injection. 

The  bacilli  are  antagonistic  to 
anthrax  bacilli.  Charrin  and  others 
have  shown  that  rabbits  inoculated 
with  a  pure- culture  of  Bacillus 
pyocyanus  after  inoculation  with 
Bacillus  an thracis  will  not  succumb 
to  anthrax.  Woodhead  and  Wood 
produced  similar  results  by  using 
sterilised  cultures,  showing  that  the 
results  were  due  to  the  chemical 
products  of  the  bacilli. 

The  rods  occur  in  the  pus  of  those 
cases  in  which  the  wounds  and  pus- 
stained  bandages  exhibit  a  greenish- 
blue  colour. 

Bacillus  pyogenes  feet  id  us 
(Passet). — Small  rods,  about  1'45  /u, 
in  length,  and  -58  p.  in  width  ; 


FIG.  209.— BACILLUS   PYOGENES 
FCETIDUS,   x  790  (PASSET). 

often  in  pairs,  or  linked  together 
in  chains.  They  are  motile,  and 
spore-formation  occurs. 

Colonies  like  white  points  appear 
after  twenty-four  hours,  and  de- 
velop into  greyish  spots,  and  these 
•enlarging  coalesce  into  a  layer. 

Cultivated  in    nutrient  gelatine 


a   greyish,  veil -like   growth   forms 
on  the  surface. 

In  nutrient  agar-agar  the  culti- 
vation resembles  the  growth  in 
gelatine.  On  blood  serum  a  moder- 
ately thick  greyish-white  streak 
develops,  and  on  sterilised  potato 
an  abundant,  shining,  brownish 
culture. 

From  all  these  media  a  putrid 
odour  emanates,  but  no  smell  is 
detected  from  a  cultivation  in  milk. 

Inoculated  into  mice  and  guinea- 
pigs,  abscesses  are  produced  or  death 
from  septicaemia  results. 

They  were  isolated  from  putrid 
pus. 

Bacillus  pyogenes  soli(Bolton). 
— Rods  resembling  Bacillus  diph- 
theriae. 

Colonies  granular,  faintly  yellow. 

The  bacilli  inoculated  in  the  depth 
of  gelatine  form  colonies  in  the 
track  of  the  needle. 

They  are  pyogenic  in  mice  and 
rabbits. 

They  are  present  in  earth. 

Bacillus  radiatus  (Luderitz).— 
Rods  4  to  7  /i  in  length,  *8  p  in 
width,  and  filaments.  Motile. 
They  are  anaerobic.  Spore-forma- 
tion present. 

Colonies  are  composed  of  delicate 
interlacing  filaments. 

In  the  depth  of  gelatine  a  growth 
occurs  at  the  lower  part  of  the 
needle  track,  from  which  fine  fila- 
ments are  given  off  in  the  sur- 
rounding gelatine,  and  liquefaction 
follows.  The  growth  in  the  depth 
of  agar  is  also  composed  of  fine 
filaments. 

In  sub-cultures  they  produce  a 
cloudy  liquefaction. 

Cultures  have  a  peculiar  odour. 

They  occur  in  earth. 

Bacillus  radiatus  aquatilis 
(Zimmermann). — Rods  1  to  6'5  /* 
in  length,  -65  //  in  width. 

Colonies  white,  with  a  marginal 
zone  of  radiating  filaments. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  grow  in  the  track 
of  the  needle,  and  excavate  and 
liquefy  the  surrounding  gelatine, 
and  form  on  the  free  surface  a 
wrinkled  patch,  which  later  subsides 


DESCRIPTION    OF   SPECIES. 


531 


to  the  bottom  of  the  liquefied 
area. 

On  agar  a  smooth  slightly 
brownish  layer  is  formed,  and  on 
potato  it  is  yellowish. 

They  occur  in  water. 

Bacillus  ramosus  (Eisenberg).— 
Rods  singly,  and  in  chains  ;  fila- 
ments. Spore -formation  present. 

Colonies  are  composed  of  curi- 
ously twisted  filaments. 

The  bacilli  inoculated  in  the  depth 
of  gelatine  produce  delicate  fila- 
ments extending  in  all  directions 
from  the  track  of  the  needle,  fol- 
lowed by  liquefaction  ;  later  a  skin 
forms  on  the  surface.  There  is  a 
sediment  at  the  bottom  of  the  tube. 

On  agar  they  form  a  greyish 
filamentous  layer,  and  on  potato 
a  whitish  growth. 

They  occur  in  earth  and  water. 

Bacillus  reticularis  (Jordan).— 
Rods  5  p,  in  length.  1  /x  in  width  ; 
singly,  and  in  short  chains.  Motile. 

Colonies  are  composed  of  radi- 
ating filaments,  and  liquefy  the 
gelatine,  forming  an  excavation  with 
a  reticulated  lining. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  filaments 
extending  from  the  track  of  the 
needle,  and  at  the  upper  part  the 
jelly  is  excavated  in  the  form  of 
a  cup.  On  agar  they  form  a  dry 
layer,  and  on  potato  a  white  woolly 
growth. 

Broth  is  made  turbid,  and  milk 
slowly  coagulated. 

They  occur  in  water. 

Bacillus  rosaceus  metalloides 
( Bacterium  rosaceum  metalloides, 
Dowdeswell  ;  Magenta  bacillus). — 
Rods  -6  to  -8  \i  in  breadth. 

Colonies  in  the  depth  of  gelatine 
are  colourless,  but  superficial  ones 
are  prominent  and  magenta  in 
colour. 

On  the  surface  of  obliquely  solidi- 
fied gelatine  they  form  a  beautiful 
magenta  band  with  a  metallic 
lustre.  The  gelatine  is  not  lique- 
fied. Similar  growths  are  obtained 
on  agar  and  potato. 

It  is  one  of  the  most  striking 
of  all  the  chromogenic  bacteria. 
Cultures  have  the  appearance  of 


having  been  stained  with  an  alco- 
holic solution  of  fuchsine.  The 
colour  varies  in  subcultures  from 
magenta  to  a  sealing-wax-red. 

Bacillus  rubefaciens  (Zirnmer- 
mann). — Rods  '75  to  1/65  \L  in 
length,  ~3'2  p.  in  width,  singly,  in 
pairs,  and  in  chains. 

Colonies  are  faintly  reddish- 
yellow. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  grow  along  the 
track  of  the  needle  and  form  a 
greyish  layer  on  the  surface  :  later 
the  jelly  acquires  a  reddish  tint. 

On  agar  the  growth  is  grey  and 
abundant. 

On  potato  the  growth  is  at  first 
grey,  later  reddish-brown,  and  the 
surface  of  the  potato  has  a  pink 
discoloration. 

They  occur  in  water. 

Bacillus  rubellus  (Okada). — 
Rods  resembling  those  of  malignant 
oedema.  They  occur  singly,  in 
pairs,  and  filaments  ;  are  motile, 
and  possess  flagella,  and  are  often 
capsulated.  They  are  anaerobic. 

Colonies  are  whitish,  with  off- 
shoots in  the  surrounding  gelatine, 
which,  later,  is  liquefied,  and  has  a 
reddish  tinge. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  growth 
in  the  lower  part  of  the  needle 
track  composed  of  isolated  colonies 
with  radiating  processes.  The  jelly 
is  liquefied,  at  first  in  the  part 
corresponding  with  the  growth,  and 
later  completely.  The  liquefied 
gelatine  is  coloured  red. 

In  agar  the  growth  extends  from 
below  upwards,  and  the  jelly  is 
coloured  red. 

In  broth  they  grow  rapidly. 

They  were  isolated  from  dust. 

Bacillus  ruber  (Breunig,  Bacille 
rouye  de  Kiel.  Laurent). — Rods 
2*5  to  5  p.  long,  and  •?  to  "8/n  broad. 
Slightly  motile. 

Colonies  below  the  surface  of 
gelatine  are  pale  yellow,  and  super- 
ficial ones  are  blood -red. 

Inoculated  in  the  depth  of  gela- 
tine they  liquefy  it  and  colour  it 
bright  red.  There  is  also  forma- 
tion of  gas  bubbles. 


532 


DESCRIPTION   OF   SPECIES. 


Potato  is  rapidly  covered  with  a 
purplish-red  growth.  Broth  be- 
comes turbid,  and  pink  in  colour. 

Milk  is  coagulated,  and  a  blood- 
red  colour  develops  on  the  surface 
and  gradually  extends. 

They  were  found  in  water. 

Bacillus  rubescens  (Jordan).— 
Rods  4  p  in  length,  -9  p,  in  width, 
singly,  in  pairs,  and  short  chains. 

Colonies  pure-white. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  little  growth  in  the 
track  of  the  needle,  and  a  pure- 
white  prominent  patch  on  the  free 
surface. 

On  agar  the  growth  is  white  and 
shining,  and  later  has  a  pink  tinge. 

On  potato  the  growth  is  flesh- 
coloured. 

Broth  becomes  turbid,  and  a  scum 
forms  on  the  surface. 

Milk  after  a  time  acquires  a  pink- 
ish colour. 

They  occur  in  sewage. 

Bacillus  rubidus  (Eisenberg).— 
Rods  and  filaments. 

Colonies  circular,  granular,  and 
slightly  red. 

The  bacilli  inoculated  in  the  depth 
of  gelatine  produce  liquefaction  and 
a  brownish-red  colour. 

On  agar  and  potato  they  form  a 
brownish-red  growth,  and  liquefy 
blood  serum. 

They  occur  in  water. 

Bacillus  sanguinis  t  y  p  h  i 
(Brannan  and  Cheeseman).— Rods 
1  to  2'5  p  in  length,  -5  to  '8  \L  in 
width ;  singly,  in  pairs,  and  in 
chains,  and  involution  forms. 

Colonies  granular,  pale-brown. 

The  bacilli  inoculated  in  the 
depth  of  glycerine-agar  produce  a 
growth  in  the  track  of  the  needle 
composed  of  isolated,  minute,  white 
colonies. 

Ra'bbits  inoculated  die  in  from 
two  weeks  to  a  month. 

They  were  isolated  from  the 
blood  of  patients  suffering  from 
typhus  fever. 

Bacillus  saprogenes  (Rosen- 
bach). — Three  rod-formed  organ- 
isms have  been  described  by 
Rosenbach  as  intimately  associ- 
ated with  putrefactive  processes. 


No.  1.— Large  rods  (Fig.  210), 
which  form  an  irregular  sinuous 
streak  with  a  mucilaginous  appear- 
ance when  cultivated  on  nutrient 
agar-agar.  Spore-formation  pre- 
sent. They  grow  also  very  readily 
on  blood  serum,  and  all  cultivations 
yield  the  odour  of  rotting  kitchen 
refuse.  They  are  not  pathogenic. 

No.  2. — Rods  shorter  and  thinner 
than  No.  1.  They  develop  very 
rapidly  on  agar-agar,  forming  trans- 
parent drops,  which  become  grey. 
The  cultivations  yield  a  character- 
istic odour  similar  to  the  last. 


FIG.  210.— BACILLUS  SAPROGENES,  No.  1. 
(Rosenbach. ) 

They  are  pathogenic  in  rabbits. 
They  appear  to  be  identical  with 
Bacillus  fcetidus  (Bacterium  fceti- 
dum,  Thin).  They  were  isolated 
from  a  patient  suffering  from  pro- 
fusely-sweating feet. 

No.  3. — See  Bacterium  sapro- 
genes. 

Bacillus  scissus  (Frankland).— 
Very  short  rods,  1  to  2  p  in  length, 
and  1  p.  in  width. 

Colonies  yellowish,  opaque  in  the 
centre,  and  periphery  dentated. 

Inoculated  in  the  depth  of  gela- 
tine there  is  no  growth  in  the 
track  of  the  needle,  but  a  shining 
layer  forms  on  the  surface,  and  the 
jelly  is  coloured  greenish. 

On  agar  the  growth  is  shining 
and  the  jelly  coloured  green. 

On  potato  the  growth  is  flesh- 
coloured. 

They  occur  in  earth. 

Bacillus  septicaemias  haemor- 
rhagicae  (p.  231). 

Bacillus  septicus  (Klein).— 
Rods  varying  in  size.  Non-motile. 
They  form  threads  or  leptothrix 
filaments,  and  are  rounded  at  the 
ends.  They  are  anaerobic,  and  form 
spores  independently  of  access  to  air. 


DESCRIPTION   OF  SPECIES. 


533 


In  a  nourishing  fluid  they  are 
overcome  by  the  presence  of  micro- 
cocci,  Bacterium  termo  or  Bacillus 
subtilis. 

They  occur  in  the  soil,  in  putrid 
blood,  and  many  putrid  albuminous 
fluids,  and  occasionally  in  the  blood- 
vessels of  man  and  animals  after 
death. 

Bacillus  septicus  acuminatus 
(Babes). — Rods  with  lancet-shaped 
ends,  about  the  size  of  the  bacilli 
of  mouse-septicaemia.  They  exhibit 
polar  staining.  They  can  be  culti- 
vated at  37°  C. 

On  agar  and  blood  serum  the 
colonies  are  circular,  transparent, 
and  later  coalesce  and  form  a  yel- 
lowish layer. 

They  are  fatal  to  rabbits  and 
guinea-pigs  in  from  two  to  six 
days. 

They  were  isolated  from  an  infant 
after  death  from  septic  infection 
occurring  five  days  after  birth. 

Bacillus  septicus  agrigenus 
(Nicolaier). — Rods  resembling  Ba- 
cillus septicaemias  haemorrhagicse. 

Colonies  circular,  granular,  with 
concentric  zones  of  varying  tints  of 
brown. 

Intravenous  injections  are  fatal 
to  rabbits  in  twenty-four  to  thirty- 
six  hours,  and  bacilli  abound  in  the 
blood. 

They  occur  in  recently  manured 
soil. 

Bacillus  septicus  keratoma- 
laciae  (Babes). — Short  thick  rods 
singly,  and  in  pairs  ;  often  capsu- 
lated. 

Colonies  white,  with  dentated 
contours. 

The  bacilli,  inoculated  in  the  depth 
of  gelatine,  grow  in  the  track  of 
the  needle  and  on  the  surface  ;  gas 
bubbles  are  developed. 

On  agar  the  growth  is  arbores- 
cent and  opalescent. 

On  blood  serum  they  form  a 
shining,  somewhat  transparent, 
dentated  film.  Cultures  have  an 
ammoniacal  odour. 

They  produce  purulent  inflam- 
mation of  the  cornea. 

They  were  isolated  from  the 
cornea  in  a  case  of  septicaemia 


following  keratomalacia  in  a 
child. 

Bacillus  septicus  ulceris  gan- 
graenosi  (Babes). — Short  rods  -5  to 
•6  p.  in  width. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  and 
gas  in  the  track  of  the  needle. 

On  agar  greyish-white  shining 
patches  are  found. 

On  potato  they  develop  a  trans- 
parent film. 

They  are  pyogenic  in  rabbits  and 
mice. 

They  were  isolated  from  the 
internal  organs  and  blood  in  a  case 
of  septicaemia  following  gangrene. 

Bacillus  septicus  vesicae 
(Clado).— Rods  1-6  to  2  /t  in  length, 
•5  p.  in  width. 

Colonies  circular,  transparent, 
yellowish. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  form  a  delicate 
filament  composed  of  closely-packed 
colonies,  and  on  the  surface  there  is 
a  filmy  growth. 

On  agar  they  form  a  greyish- 
white  layer,  and  on  potato  the 
growth  is  dry  and  brown. 

They  are  poisonous  to  rabbits, 
guinea-pigs,  and  mice. 

They  were  isolated  from  urine 
from  a  case  of  cystitis. 

Bacillus  sessilis  (L.  Klein).— 
Rods  resembling  those  of  the  hay 
bacillus. 

They  are  said  to  be  distinguished 
by  fission  commencing  in  a  newly 
formed  rod  before  it  has  been  set 
free  from  the  spore. 

They  were  isolated  from  the  blood 
of  a  cow. 

Bacillus  smaragdino-phosplio- 
rescens  (Katz). — Rods  with  pointed 
ends.  2  p  in  length,  1  /LI  in  width. 

Colonies  circular,  faintly  yellow, 
with  concentric  rings. 

Inoculated  in  the  depth  of  gela- 
tine a  white  filament  forms  in  the 
track  of  the  needle  and  a  greyish- 
white  patch  on  the  free  surface  ; 
and  there  is  sometimes  liquefaction. 

In  broth  they  produce  turbidity. 

On  potato  they  produce  a  thin 
brownish-yellow  film. 

They  are    photogenic,   and    the 


534 


DESCRIFIION   OF   SPECIES. 


phosphorescence  is  most  marked  in 
cultures  containing  an  excess  of 
salt. 

They  were  isolated  from  a  phos- 
phorescent herring. 

Bacillus  smaragdinus  fcetidus 
(Reimann). — Slender  rods  slightly 
bent. 

Colonies  on  agar  irregular,  with 
a  yellowish  granular  nucleus,  and 
transparent  marginal  zone. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  growth 
in  the  track  of  the  needle,  and 
liquefaction  at  its  upper  part, 
and  a  greenish  coloration. 

In  the  depth  of  agar  the  medium 
is  coloured  green. 

On  potato  the  growth  is  brown. 
Cultures  emit  a  strong  odour. 

Intravenous  injection  produces 
death  in  rabbits  in  forty-eight 
hours. 

They  were  isolated  from  nasal 
mucus  in  ozsena. 

Bacillus  solidus  (Liideritz).— 
Rods  1  to  10  /A  in  length,  -5  p.  in 
width. 

They  are  anaerobic.  Cultivated  in 
grape-sugar  gelatine  they  produce 
gas  bubbles  and  a  penetrating  foul 
odour.  The  colonies  are  spherical, 
and  in  agar  under  a  low  power 
are  seen  to  be  composed  of  fine 
filaments  like  cotton  wool. 

In  broth  with  exclusion  of  oxygen 
they  produce  a  copious  growth  with 
abundant  formation  of  foetid  gas. 
They  were  isolated  from  earth. 
Bacillus  spiniferus  (Unna).— 
Rods   sometimes    curved,   2    p.    in 
length,  -8  to  1  p  in  width,  singly, 
in  pairs,  and  masses. 

Colonies  have  peculiar  spines,  and 
later  a  radiated  marginal  zone. 

In  the  depth  of  gelatine  minute, 
yellowish,  isolated  colonies  develop 
in  the  track  of  the  needle,  and  a 
furrowed,  yellowish-grey  patch  on 
the  free  surface. 

On  agar  the  same  yellowish 
wrinkled  growth  appears. 

On  potato  they  form  a  shining, 
faintly-yellow  layer. 

They  were  isolated  from  the  skin 


Rods  sometimes  bent,  3  to  8  p.  in 
length,  -6  n  in  width,  and  long 
filaments.  Spore-formation  present. 
They  are  anaerobic. 

Colonies  are    composed   of    fine 
radiating  filaments,  and  liquefy  the 


in  eczema. 


Bacillus  spinosus  (Luderitz). — 


In  agar  the  growth  is  composed 
of    colonies    of    matted   filaments, 
and  there  is  gas-formation. 
They  liquefy  blood  serum. 
They  occur  in  earth. 
Bacillus  stolonatus  (Adametz), 
— Rods  motile. 

Colonies  on  gelatine,  circular, 
granular,  and  whitish,  or  yellowish- 
brown.  Colonies  on  agar  send  off 
peculiar  wavy  processes. 

Inoculated  in  the  depth  of  gela- 
tine a  granular  filament  develops  in 
the  track  of  the  needle,  and  a  white 
patch  on  the  free  surface  ;  later  the 
gelatine  is  excavated  in  the  upper 
part,  and  the  culture  lines  the 
cavity. 

On  potato  the  growth  is  whitish. 
They  occur  in  water. 
Bacillus  stoloniferus  (Pohl).— 
Rods  1'2  p.  in  length,  '8  /*  in  width. 
Motile. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  rapid  liquefaction 
!  in  the  track  of  the  needle. 

On  the  surface  of  agar  they  form 
;  a  white  growth. 

On  potato  they  grow  abundantly, 
I  but  scarcely  at  all  in  milk. 

They  occur  in  the  water  of 
marshes. 

Bacillus  striatus  albus  (Bes- 
ser). — Rods  sometimes  bent. 

Colonies  on  gelatine  appear  as 
minute  dry  points. 

On  agar  the  colonies  have  a  brown 
1   nucleus  and  clear  marginal  zone. 

On  the  surface  of  agar  the  bacilli 
produce  a  greyish- white  thin  layer. 
On  potato  the  growth  is  trans- 
parent and  slightly  gelatinous. 
They  occur  in  nasal  mucus. 
Bacillus  striatus  flavus  (Bes- 
ser). — Short      rods,      straight     or 
curved  ;  involution  forms. 
Colonies  granular,  yellowish. 
On  the  surface  of  agar  they  pro- 
duce a  white  growth,  which  later 
becomes  sulphur  yellow. 


DESCRIFHON    OF   SPECIES. 


535 


On  potato  a  similar  colour  is 
produced. 

They  were  isolated  from  nasal 
mucus. 

Bacillus  subflavus  (Zimmer- 
mann). — Rods  1-5  to  3  p.  in  length, 
'11  fj.  in  width,  and  in  chains.  Motile. 

Colonies  prominent,  yellowish- 
white. 

On  the  surface  of  gelatine  they 
form  a  yellowish-grey  layer,  and 
on  the  surface  of  agar  and  potato 
the  growth  is  yellow. 

They  occur  in  water. 

Bacillus  subtilis  (Hay  bacillus). 
— Cylindrical  rods  as  much  as  6  /x 
in  length.  Single  forms  grow  to 
double  their  length,  and  then 
undergo  division.  They  also  form 
threads  which  may  be  composed  of 


FIG. 


211. — BACILLUS    SUBTILIS 
SPORES  (BAUMGARTEN). 


long  rods,  short  rods,  and  cocci. 
They  are  motile,  and  provided  with 
a  flagellum  at  each  end.  If  the 
nourishing  medium  is  impoverished, 
the  multiplication  of  the  rods  by 
division  gradually  ceases,  and  spore- 
formation  commences.  The  rods 
become  motionless,  and  a  dark  spot 
is  visible,  either  in  the  middle  or 
towards  one  end.  This  gradually 
develops  into  a  shining  spore  with 
a  dark  outline.  The  rods  swell 
slightly  during  this  process  ;  their 
contour  becomes  undefined,  and 
soon  disappears  entirely  ;  spores 
being  set  free  in  about  twenty- 
four  hours.  The  spores  are  1'2  p. 
long,  and  *G  /x  broad.  They  develop 
into  rods  in  the  following  way  : — 
On  one  side  of  the  spore  a  swelling 
appears,  at  the  summit  of  which 
an  opening  in  the  spore-membrane 
results,  and  the  germ  escapes  ;  this 
lengthens  into  a  rod,  and  remains 
for  a  time  attached  to  the  empty 
spore-membrane. 


The  spores  are  widely  distributed, 
and  occur  in  the  air,  soil,  dust,  etc. 
On  the  excrement  of  herbivorous 
animals  the  bacilli  form  a  white 
efflorescence,  and  a  thick  crumpled 
skin  on  liquid  manure. 

They  flourish  equally  in  liquids 
and  upon  damp,  solid,  nourishing 
media.  They  are  aerobic  ;  depriva- 
tion of  oxygen  causes  the  growth 
of  the  bacilli  to  cease,  and  the  rods 
degenerate. 

In  plate-cultivations  the  colonies 
are  white,  and,  under  a  low  power, 
granular  and  irregular  in  outline 
and  faintly-greenish.  Liquefaction 
sets  in,  producing  depressions  like 
saucers.  The  centre  is  opaque,  and 
is  surrounded  by  a  network  of 
filaments,  which  extend  into  the 
gelatine  surrounding  the  colony. 


FIG.  212. — PURE-CULTURE  OF  BACILLI'S 
SUBTILIS  IN  NUTRIENT  GELATINE 
(BAUMGARTEN). 

Inoculated  in  the  depth  of  gela- 
tine, liquefaction  occurs  rapidly  in 
the  track  of  the  needle,  and  a  film 


536 


DESCRIPTION   OF   SPECIES. 


floats  on  the  surface.  The  liquefied 
gelatine,  at  first  turbid,  becomes 
clear  as  the  bacilli  settle  at  the 
bottom  of  the  tube. 

On  agar  a  wrinkled  film  develops, 
and  also  on  serum. 


FIG.  213.    PURE-CULTURE  OF  BACILLUS 

SUBTILIS  ON   THE   SURFACE  OF   Nu- 

TRIENT  AGAR. 

On  potato  the  growth  is  white, 
and  there  is  copious  spore-forma- 
tion. 

On  ordinary  nutrient  liquids  they 
develop  at  first  a  thin,  and  subse- 
quently a  thick,  dense,  crumpled 
pellicle,  with  copious  spore-forma- 
tion. 

The  simplest  way  to  obtain  a 
culture  of  the  bacillus  is  to  make 
a  decoction  of  hay.  The  hay  is 
chopped  into  small  pieces,  and 
boiled  with  distilled  water  in  a 
flask  for  a  quarter  of  an  hour. 
The  infusion  is  then  filtered  into  a 
beaker,  covered  with  a  glass  plate, 
and  set  aside  in  a  warm  place.  In 
two  or  three  days,  the  liquid  swarms 


with  the  bacilli,  the  spores  of  which 
exist  in  great  numbers  in  ordinary 
hay.  A  more  sure  method  for 
obtaining  a  pure  cultivation  is  as 
follows  : — 

(«)  Add  only  a  small  quantity  of 
water  to  some  finely  chopped  hay, 
and  set  aside  for  four  hours  at 
36°  C. 

(6)  Pour  off  the  extract,  and 
dilute  it  to  the  sp.  gr.  1'004. 

(c)  Boil  gently  for  one  hour  in 
a  bulb  plugged  with  cotton  wool. 

(d)  Set   aside   500   ccm.  of  the 
extract  at  36°  C. 

In  about  twenty-four  hours,  as 
a  rule,  a  pellicle  has  commenced 
to  develop  upon  the  surface  of  the 
liquid.  If  the  reaction  is  definitely 
acid,  carbonate  of  soda  solution 
must  be  added  to  the  decoction. 

METHODS  OF  STAINING  HAY  BACILLUS. 

To  demonstrate  the  flagella  of  the 
bacilli,  they  may  be  stained  with 
hsematoxylin  solution  (Koch),  or  by 
Loffler's  method. 

The  linking  together  of  cocci,  long 
rods  and  short  rods  in  the  threads,  is 
shown  by  treating  with  alcoholic  solu- 
tion or  f  uchsine,  or  with  iodine  solution 
(Zopf). 

To  stain  the  spores  the  cover-glass 
preparations  must  be  heated  to  a  very 
high  temperature  (210UC.),  in  the  hot- 
air  steriliser  for  half  an  hour,  or  they 
may  be  exposed  for  a  few  seconds  to 
the  action  of  concentrated  sulphuric 
acid  (Biichner),  or  floated  for  twenty 
minutes  on  hot  solution  of  the  dye. 

Bacillus  subtilis  similans.— 
There  are  several  bacilli  closely 
resembling  Bacillus  subtilis. 

Two  have  been  isolated  from 
human  faeces  by  Bienstock  which 
do  not  liquefy  nutrient  gelatine. 

No.  I.  Rods  and  filaments ; 
spore-formation  present. 

On  agar  they  produce  a  delicate 
wrinkled  veil. 

No.  II.  Rods  morphologically 
identical  with  No.  I. 

On  agar  they  produce  a  smooth, 
shining  layer. 

Bacillus  superficialis  (Jordan). 
— Rods  2 '2  p.  in  length,  and  '1  p,  in 
width  ;  singly,  and  in  pairs.  Motile. 

Colonies  have  a  yellowish-brown 


DESCRIPTION    OF   SPECIES. 


537 


nucleus  and  transparent  marginal 
zone. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  slight  growth  in  the 
track  of  the  needle,  and  after  a  time 
liquefaction  at  the  upper  part. 

On  agar  the  growth  is  smooth  and 
shining. 

In  broth  they  produce  turbidity. 

They  will  not  grow  on  potato. 

They  occur  in  sewage. 

Bacillus  tennis  sputigenus 
(Pansini). — Short  rods,  singly,  and 
in  pairs  ;  capsulated. 

They  produce  a  whitish  growth 
on  the  surface  of  gelatine. 

They  coagulate  milk. 

They  are  pathogenic  in  rabbits. 

They  were  isolated  from  sputum. 

Bacillus  termo  (Mace).— Thick 
rods  1*4  \t.  long,  and  '8  p.  wide, 
usually  in  pairs,  sometimes  in  chains. 
Actively  motile. 

Colonies  whitish,  with  a  grey  edge 
surrounded  by  liquefied  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  funnel-shaped 
area  of  liquefaction,  and  later  the 
whole  of  the  jelly  is  liquefied. 

Broth  is  rendered  turbid  and  a 
thin  brittle  pellicle  is  formed. 

They  are  associated  with  decom- 
position. 

Bacillus  tetani  (p.  457). 

Bacillus  thalassophilus  (Rus- 
sel). — Slender  rods  varying  in 
length  ;  and  filaments.  They  are 
anaerobic.  Spore-formation  pre- 
sent. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  appears  in  the 
lower  part  of  the  track  of  the 
needle  in  the  form  of  cloudy  colo- 
nies, liquefying  the  jelly  and  pro- 
ducing gas-bubbles.  Cultures  emit 
a  penetrating  odour. 

They  were  isolated  from  sea-mud. 

Bacillus  thermophilus  (Mi- 
quel). — Rods  vary  ing  in  size  accord- 
ing to  the  temperature  at  which 
they  are  cultivated.  In  broth  they 
grow  best  between  65°  and  70°  C., 
forming  a  copious  deposit.  They 
occur  in  air,  soil,  and  water. 

Bacillus  tremelloid.es  (Tils).— 
Rods  -75  to  1  /i  in  length,  -25  p  in 
width  ;  and  in  masses. 


Colonies  circular,  yellowish- 
brown. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  growth 
composed  of  isolated  yellow  colo- 
nies in  the  track  of  the  needle, 
and  a  yellow  mass  on  the  surface. 
They  liquefy  the  gelatine. 

On  agar  the  growth  is  slimy  and 
golden-yellow. 

On  potato  they  form  an  abund- 
|   ant  yellow  growth. 
They  occur  in  water. 
Bacillus  tuberculosis  (p.  378). 
Bacillus     tuberculosis    galli- 
narum  (p.  402). 

Bacillus    tumescens   (Zopf). — 
Cocci,  long  and  short  rods.     They 
i  form  a  jelly-like  disc  *5  to  1  cm.  in 
diam.   on   slices  of    boiled    carrot, 
with   the   appearance   of  a  rather 
tough,  crumpled  skin  of  a  whitish 
I   colour.     Examination  of   this   pel- 
j   licle   shows   that  it   is   formed  of 
rows  of  rods  lying  closely  together. 
These    rods    can    be    observed  to 
divide   into   short  rods  and  cocci. 
j   Spore-formation     occurs     in    two 
stages  of  development — viz.,  in  the 
|   cocci   and   in   the   short  rods.      A 
j   cultivation  is  obtained  by  exposing 
'   slices    of    boiled     carrot,    slightly 
!   moistened,  to  the  air  at  the  tem- 
perature of  the  room. 

Bacillus  typhi  abdominalis 
(p.  342). 

Bacillus  ubiquitus  (Jordan).— 
Rods   1*1    to  2  p.   in   length,  '1   /A 
I  in  width  ;  and  filaments. 

Colonies    -granular      and     well 
I  defined. 

The    bacilli    inoculated    in    the 
|  depth  of  gelatine  produce  a  growth 
!  resembling   that    of    Friedlanders 
pneumococcus. 

On  agar  and  potato  the  growth 
i  is  greyish-white. 

They  coagulate  milk  and  reduce 
nitrates. 

They  occur  in  air  and  water. 
Probably   a   variety  of  Bacillus 
candicans. 

Bacillus  ulna  (Cohn).— Cocci, 
short  rods,  long  rods,  and  threads. 
Diam.  of  the  cocci  1*5  to  2'2  p.. 
Spore-formation  in  both  short  and 
long  rods.  No  septic  odour  is  pro- 


538 


DESCRIPTION   OF   SPECIES. 


duced  by  this  bacillus  in  a  nourish- 
ing liquid.  Cloudy  masses  are 
found  on  the  surface  of  the  liquid, 
which  later  form  a  thick  dry 
pellicle,  consisting  of  bundles  of 
threads  matted  together.  The  for- 
mation of  ellipsoidal  spores  occurs 
in  the  usual  way ;  they  measure 
2  '5  to  2 '8  \i  long,  and  more  than 

1  /A  wide.     The  bacillus  is  found 
in  rotting  eggs,  and  can  be  culti- 
vated on  boiled  white  of  egg. 

Bacillus  ulna  (Vignaf).— Rods 

2  \L  in  length  ;  singly,  and  in  pairs, 
and  in  short  chains. 

Colonies  composed  of  concentric 
zones  varying  in  granularity. 

Inoculated  in  the  depth  of  gela- 
tine, liquefaction  occurs  rapidly  in 
the  track  of  the  needle  ;  later,  there 
is  a  deposit  at  the  bottom  of  the 
liquefied  area  and  ji  pellicle  on  the 
surface. 

On  agar  they  form  a  white  ad^ 
herent  layer,  and  the  jelly  is  tinged 
with  brown. 

In  broth  a  pellicle  forms  on  the 
surface. 

On  potato  they  form  a  pellicle 
with  characteristic  linear  markings. 

They  liquefy  serum.  Cultures 
produce  a  putrefactive  odour. 

They  occur  in  human  saliva. 

Bacillus  vacuolpsis  (Sternberg). 
— Rods  1%5  to  5  p.  in  length,  1  /A  in 
width,  containing  vacuolated  proto- 
plasm; filaments,  and  involution 
forms.  At  times  slowly  motile. 

Inoculated  in  the  depth  of  gela- 
tine, liquefaction  occurs  slowly  at 
the  upper  part  of  the  track  of 
the  needle,  forming  a  cup-shaped 
cavity  ;  the  liquefied  gelatine  is 
viscid,  and  a  cream-white  layer 
forms  on  the  surface. 

In  agar  the  development  in  the 
track  of  the  needle  is  scanty  ;  on 
the  surface  a  cream-white  layer  is 
formed,  and  the  bacilli  are  united 
in  long  jointed  filaments. 

On  potato  a  similar  growth  is 
produced. 

They  were  isolated  from  the  in- 
testine in  fatal  cases  of  yellow  fever. 

Bacillus  varicosus  conjunctivae 
(Gombert).— Rods  2  to  8  /*  m  length, 
1  p.  in  width. 


Inoculated  in  the  depth  of  gela- 
tine they  produce  a  greyish-white 
filament  in  the  track  of  the  needle, 
and  a  greyish-white  patch  on  the 
surface  ;  liquefaction  follows  with- 
out turbidity. 

On  the  surface  of  agar  a  white, 
dry.  adherent  film  is  formed. 

On  potato  the  growth  is,  at  first, 
white  and  dry,  later,  reddish-brown. 

They  produce  hypersemia  when 
injected  into  the  conjunctiva. 

They  were  isolated  from  the 
healthy  human  conjunctiva. 

Bacillus  venenosus  (Vaughan). 
— Motile  rods. 

Colonies  circular,  whitish. 

Inoculated  in  the  depth  of  gela- 
tine there  is  growth  in  the  track  of 
the  needle  and  on  the  free  surface. 

On  agar  they  form  a  white  film, 
and  on  potato  a  moist  brownish 
layer. 

They  are  pathogenic  in  small 
animals. 

They  occur  in  water. 

Bacillus  venenosus  brevis 
(Vaughan). — Rods  short  and  thick. 

Colonies  are  yellow  and  composed 
of  concentric  rings. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  in  the  track  of  the 
needle  and  over  the  free  surface. 

On  agar  they  produce  a  white 
film. 

On  potato  the  growth  is  brownish. 

They  are  pathogenic  in  small 
animals. 

They  occur  in  water. 

Bacillus  venenosus  invisibilis. 
— Slender  rods. 

Colonies  irregular,  granular. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  extremely  slow 
both  in  the  track  of  the  needle  and 
on  the  surface. 

On  agar  there  is  a  whitish  film, 
and  on  potatoes  a  brownish  layer. 

They  are  pathogenic  in  small 
animals. 

They  occur  in  water. 

Bacillus  venenosus  lique- 
faciens  (Vaughan).— Rods. 

CoJonies  circular,  granular,  yel- 
lowish. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  in  the  track  of 


DESCRIPTION    OF   SPECIES. 


539 


the  needle  and  on  the  surface,  and 
liquefaction  occurs  after  some 
weeks. 

On  agar  they  produce  a  white 
growth,  and  on  potato  it  is  brown- 
ish or  yellowish. 

They  are  pathogenic  in  small 
animals. 

They  occur  in  water. 

Bacillus  ventriculi  (Raczyn- 
sky). — Rods  1-5  to  3  p  in  length, 
1  /z  in  width,  singly,  in  pairs,  and 
in  short  chains. 

Colonies  have  a  dark  nucleus  and 
transparent  periphery. 

On  agar  they  form  a  white  layer. 

They  were  isolated  from  the 
digestive  tract  of  dogs. 

Bacillus  vermicularis  (Frank- 
land). — Large  bacilli  2  to  3  /*  in 
length,  1  /z  in  width,  and  long 
threads.  Spore-formation  present. 

Colonies  are  irregular  in  contour, 
the  irregularity  increasing  as  the 
colony  comes  to  the  surface.  The 
peripheral  part  is  composed  of 
closely  packed,  wavy  bands  of  bacilli, 
and  the  centre  is  irregular  and 
wrinkled. 

The  bacilli  inoculated  in.  the 
depth  of  gelatine  form  a  flattened 
band  in  the  track  of  the  needle,  and 
a  grey  layer  on  the  surface  ;  lique- 
faction slowly  follows. 

On  agar  they  produce  a  smooth,, 
shining,  grey  layer,  and  on  potato 
a  thick,  irregular,  flesh-coloured 
growth. 

They  reduce  nitrates. 

They  occur  in  water.  Probably 
identical  with  Bacillus  vermicu- 
losus. 

Bacillus  vermiculosus  (Zimmer- 
mann). — Rods  1'5  /z  in  length,  -85  p 
in  width,  singly,  in  pairs,  very  short 
chains  and  long  filaments.  They 
are  slowly  motile. 

Colonies  irregular  ;  grey,  granu- 
lar. 

Inoculated  in  the  depth  of  gela- 
tine they  produce,  after  four  days, 
liquefaction  in  the  upper  part  of 
the  needle  track,  which  spreads 
downwards,  and  a  reddish-grey  sedi- 
ment collects  at  the  bottom  of  the 
liquefied  area. 

On  agar  the  growth  is  smooth  and 


shining,  and  on  potato  yellowish- 
grey. 

They  occur  in  water. 

Bacillus  violaceus  (vide  Bacil- 
lus ianthinus). 

Bacillus  violaceus  Laurentius 
(Jordan).— Rods  3  to  3'6  p,  in  length, 
'7  p  in  width. 

Colonies  violet,  surrounded  by 
liquefied  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the  track 
of  the  needle,  and  a  violet  sediment 
collects  at  the  bottom. 

On  agar  the  growth  is  violet,  later 
black. 

On  potato  there  is  a  copious 
growth,  changing  in  colour  from 
violet  to  black. 

In  broth  a  violet  colour  is  pro- 
duced in  the  presence  of  nitrates. 

They  coagulate  milk,  and  render 
it  bluish-violet. 

They  occur  in  water.  Probably 
identical  with  Bacillus  ianthinus 
(Zopf). 

Bacillus  virescens  (Frick).— 
Rods  and  filaments. 

Colonies  irregular,  granular, 
green. 

On  the  surface  of  gelatine  they 
colour  the  medium  green. 

They  grow  on  agar. 

On  potato  they  form  a  brownish 
growth. 

In  broth  a  pellicle  is  formed  on 
the  surface,  and  beneath  it  the 
liquid  is  coloured  green. 

They  were  isolated  from  green 
sputum. 

Bacillus  viscosus  (Frankland). 
— Rods  1*5  to  "2  p.  in  length,  singly 
and  in  pairs. 

Colonies  granular,  with  hairlike 
processes  extending  into  the  gela- 
tine, which  is  liquefied  and  has  a 
green  colour. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  and 
a  green  fluorescence. 

On  agar  they  form  a  greenish- 
j  white  layer,  and  colour  the  jelly 
I  green. 

On  potato  the  growth  is  brown. 

Probably  identical  with  Bacillus 
fluorescens  liquefaciens. 

Bacillus    Zurnianus    (List).— 


\  8  R  A 

OF  THW 

UNIVERSITY 


o40 


DESCRIPTION    OF  SPECIES. 


Rods  1-2  to  1  -5  p  in  length,  -6  to  -8  A* 
in  width.  Colonies  greyish-white, 
viscid. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  develop  slightly 
in  the  track  of  the  needle,  and  pro- 
duce a  prominent  grape-like  growth 
on  the  free  surface. 

On  potato  the  growth  is  grey  or 
tinged  with  yellow. 

They  occur  in  water. 

Bacterium  aerogenes  (Miller). 
—Short  rods,  singly  and  in  pairs. 
Motile. 

The  colonies  are  circular,  well 
defined,  and  yellowish. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  in  the  track  of  the 
needle  is  brownish-yellow,  and  a 
flat  greyish  button  forms  on  the 
free  surface. 

On  agar  a  pulpy  layer  develops. 

On  potato  the  growth  is  pulpy 
and  yellowish-white. 

The  bacteria  possess  great  power 
of  resisting  the  effect  of  acids. 

They  were  isolated  from  the  diges- 
tive tract. 

Bacterium  brunneum 
(Schroter). — Motile  rods,  produ- 
cing a  brown  colour. 

They  were  observed  on  a  rotting 
infusion  of  maize. 

Bacterium  decalvans  (Thin).— 
Cocci,  singly  or  in  pairs,  1-6  //.  in 
length. 

They  were  observed  in  the  roots 
of  the  hair  in  cases  of  Alopecia 
areatti. 

Bacterium  fusiforme  (Warm- 
ing).—Rods  spindle-shaped,  with 
pointed  ends,  2-5  p.  long,  and  '5  to 
•8  n  thick.  They  were  described 
as  forming  a  spongy  layer  on  the 
surface  of  sea-water. 

Bacterium  gingivae  pyogenes 
(Miller).— Short  rods. 

The  colonies  are  circular  and 
rapidly  liquefy  gelatine. 

The  bacteria  inoculated  in  the 
depth  of  gelatine  produce  rapid 
liquefaction  in  the  track  of  the 
needle  and  a  white  sediment. 

On  agar  they  produce  a  moist 
white  growth. 

They  are  pyogenic  when  inocu- 
lated subcutaueously  in  small  ani- 


mals, and  cause  a  fatal  result  when 
injected  to  the  peritoneal  cavity. 

They  occur  in  the  deposit  on  the 
teeth. 

Bacterium  hyacinthi  (Wakker). 
—  Cocci  resembling  Bacterium 
termo. 

They  were  observed  in  the  yellow 
slime  of  diseased  hyacinth  bulbs. 

Bacterium  hydrosulfureum 
ponticum  (Zelinsky-).  —  Long 
motile  rods. 

On  agar  a  dark  coffee-coloured 
pigment  is  produced,  which  turns 
black  when  exposed  to  air. 

Cultures  give  off  sulphuretted 
hydrogen. 

They  were  isolated  fromdredgings 
in  the  Black  Sea. 

Bacterium  litoreum  (Warming). 
— Cocci  ellipsoidal,  2  to  6  /*  long, 
1'2  to  2-4  fj,  wide  ;  singly,  never 
as  chains  or  zoogloea. 

They  occur  in  sea-water. 

Bacterium  luteum  (List).— 
Rods  from  I'l  to  1'3  ^  long.  Non- 
motile.  The  colonies  are  slimy,  with 
orange  centres. 

Inoculated  in  the  depth  of 
gelatine  an  orange  growth  occurs, 
principally  at  the  point  of  puncture. 

Milk  is  coagulated. 

They  occur  in  water. 

Bacterium  merismopedioides 
(Zppf).— Threads  I  to  1-5  w  in 
thickness ;  these  subdivide  into 
long  rods,  short  rods,  and  finally 
into  cocci.  The  cocci  divide  first 
in  one  and  subsequently  in  two 
directions,  forming  characteristic 
groups,  which  appear  like  merismo- 
pedia.  These  groups  may  eventually 
consist  of  64  by  64  cells  or  more, 
and  ultimately  form  zoogloea.  The 
cocci  develop  again  into  rods  and 
threads. 

They  were  observed  in  water 
containing  putrefying  substances 
(River  Panke,  Berlin). 

Bacterium  navicula  (Reinke 
and  Berthold). — Cocci  spindle-form 
or  ellipsoidal,  including  motile  and 
non-motile  forms.  They  have  one 
or  more  dark  spots,  which  may  be 
coloured  blue  by  iodine. 

They  have  been  observed  in  rot- 
ting potatoes. 


DESCRIPTION   OF   SPECIES. 


541 


Bacterium  photometricum(En- 

gelmann). — Rods  slightly  reddish  in 
colour  ;  motile. 

The  movements  are  stated  to 
depend  on  light. 

Bacterium  synxanthum  (Ehren- 
berg  ;  Bacterium  xanthinum  ;  Bac- 
terium of  yellow  milk). — Cocci  '7  to 
1  /z  in  length,  and  rod-forms. 
They  produce  a  yellow  colour  in 
boiled  milk,  which  at  first  becomes 
acid,  and  then  strongly  alkaline. 
They  also  occur  on  boiled  potatoes, 
carrots,  etc.,  where  they  form  small 
lemon-yellow  masses. 

The  colouring-matter  is  soluble 
in  water,  insoluble  in  ether  and 
alcohol,  unchanged  by  alkalies,  de- 
colorised by  acids.  It  is  similar 
to  yellow  aniline  colours,  both 
spectroscopically  and  in  ordinary 
reactions. 

Bacterium  termo  (Vignal).— 
Rods  1*5  to  2  p.  in  length,  *5  to  '7  p. 
in  width. 

Colonies  white,  surrounded  by 
liquefied  gelatine. 

The  bacilli  inoculated  in  the 
depth  of  gelatine  produce  a  funnel- 
shaped  area  of  liquefaction  ;  later, 
the  jelly  is  completely  liquefied  and 
coloured  green.  Cultures  have  a 
strong  putrefactive  odour. 

In  broth  they  form  a  white 
deposit  and  colour  the  medium 
green. 

They  were  isolated  from  human 
saliva. 

Bacterium  tholoeideum  (Gess- 
ner). — Rods  similar  to  Bacillus 
lactis  aerogenes. 

Pathogenic  in  small  animals. 

They  were  isolated  from  healthy 
human  evacuations. 

Bacterium  ureas  (Cohn).— Cocci 
1*25  to  2  /z  in  diam.,  singly  or  in 
chains,  and  rods.  The  rods  split 
up  by  division  into  chains  of  cocci, 
which  after  a  time  are  set  free.  The 
cocci  increase  further  by  subdivi- 
sion, and  a  jelly-like  membrane 
develops  around  them.  Masses  of 
cocci  exist  in  the  form  of  irregular 
or  roundish  lumps.  They  are 
aerobic. 

Cultivations,  after  twenty-four 
hours,  consist  exclusively  of  rods  ; 


after  forty-eight  hours,  of  cocci 
chains  ;  and  in  fourteen  days,  of 
zoogloea  ;  the  cocci  transplanted 
into  fresh  nourishing  solution  again 
grow  into  rods.  These  observations 
point  to  the  existence  of  a  pleo- 
morphic  species,  Bacterium  urece  ; 
and  the  former  nomenclature,  Micro- 
coccus  urece,  must  be  regarded  as 
untenable. 

In  urine  they  set  up  ammoniacal 
fermentation,  converting  urea  into 
carbonate  of  ammonia.  Rods,  2  p. 
long  and  1  /z  wide,  have  been  iso- 
lated from  stale  urine  (Bacillus 
urese,  Leube),  which  also  most 
energetically  cause  the  ammoniacal 
fermentation  of  urine. 

Bacterium  ureae  (Jaksch).— 
Rods  2  /*  in  length,  1  /z  in  width. 

Colonies  on  gelatine  semi-trans- 
parent. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  form  a  delicate 
branching  growth  in  the  track  of 
the  needle. 

They  convert  urea  into  carbonate 
of  ammonia,  and  cultures  smell  of 
herring  brine. 

They  occur  in  ammoniacal  urine. 

Bacterium  yiolaceum  (Bergon- 
zini). — Rods  similar  to  Bacterium 
termo,  '6  to  1  /z  thick,  2  to  3  tt  long. 

They  occur  on  white  of  egg, 
forming  a  violet  pigment. 

Bacterium  Zopfii  (Kurth).— 
Cocci,  1  to  1'25  /z  in  diam.  ;  rods 
and  threads.  Cultivated  in  a  streak 
on  nutrient  gelatine  spread  out  on 
a  glass  slide,  a  peculiar  develop- 
ment takes  place.  In  twenty-four 
hours  after  inoculation  threads 
have  developed ;  in  forty-eight 
hours  windings  of  the  threads 
are  observed,  and  in  six  days  the 
threads  have  broken  up  into  cocci. 
They  were  observed  in  the  intestine 
of  fowls,  especially  in  the  contents 
of  the  vermiform  appendix.  In- 
oculation of  rabbits  was  followed 
by  negative  results.  Identical  with 
Bacillus  figurans  (Crookshank). 

Beggiatoa  alba  (Vauch).— Cocci,, 
rods,  spirals  and  threads  (Fig.  215). 
The  threads  are  indistinctly  articu- 
lated, actively  oscillating,  and  colour- 
less ;  their  protoplasm  contains 


o42 


.DESCRIPTION   OF   SPECIES. 


numerous  strongly  refractive  gran- 
ules consisting  of  sulphur.  They 
occur  as  greyish  or  chalk- white 
gelatinous  threads,  3  to  3 '5  /* 
thick,  in  sulphur  springs  and 
marshes. 

Beggiatqa  mirabilis  (Cohn).— 
Threads  distinguished  by  their 
breadth,  which  may  reach  30  p. 
They  are  motile,  bent  and  curled 
in  various  ways,  and  rounded  at 
the  ends.  Around  the  threads, 
isolated  cells  have  been  observed, 


families,  bound  together  by  gela- 
tinous substance.  Later  they  be- 
come larger,  globular  or  ovoid  in 
shape,  and  hollow,  containing 
watery  fluid  in  their  interior.  The 
families  reach  a  diameter  of  660  ju, 
in  which  the  cocci  form  simply  a 
peripheral  layer.  The  hollow  fami- 
lies or  vesicles  are  often  perforated, 
presenting  a  delicate  reticulated  ap- 
pearance, which  finally  may  become 
broken  up  into  irregular  structures. 
The  red  colouring-matter  can  be 


t>Hf  Z°P1?\    S1™CE88IY*   CHANGES   IN   THE   SAME  THREAD, 
thread  form  ;  b,  breaking  up  into  rod  forms ;  c,  into  cocci  (Kurth). 


macrococci,  but  spiral  forms  are  as 
yet  unknown.  The  threads  are 
filled  with  sulphur  granules.  They 
occur  in  sea-water,  forming  a  white 
gelatinous  scum  on  decomposing 
algae. 

Beggiatoa  r oseo-p  e  r  s  i  c  i  n  a 
{Co/mm  roseo-perttdna.  Bacterium 
rubescens,  or  Peach-coloured  bac- 
tfirtum,  Lankester).— Cocci,  rods, 
spirals,  and  threads  (Fig.  216).  The 
«occi,  globular  or  oval,  reach  2-5  /* 
an  diam.  They  form  at  first  solid 


distinguished  from  other  red  pig- 
ments, and  it  is  designated  by  the 
name  bacterio-purpurin.  It  is  quite 
distinct  from  the  pigment  produced 
by  Micrococcus  prodigiosus,  being 
peach-blossom  red,  and  insoluble 
m  water,  alcohol,  etc.  Examined 
spectroscopically,  it  shows  a  strong 
absorption  in  the  yellow,  and  a 
weaker  band  in  the  green  and  blue, 
as  well  as  a  darkening  in  the  more 
refrangible  half  of  the  spectrum. 
In  the  cocci,  especially  of  the  older 


DESCRIPTION   OF   SPECIES. 


543 


vesicles,  dark  granules  are  to  be 
seen,  which  consist  of  sulphur. 
The  micro-organisms  occur  on  the 
surface  of  marshes,  or  on  water  in 
which-- 1  algge  are  rotting.  They 
form  a  rose-red,  blood-red,  violet- 
red,  or  violet-brown  scum  ;  and 
sometimes  in  such  quantity  that 


Cladothrix  dichotoma  (Cohn). 
— Threads  resembling  those  of  lep- 
tothrix ;  slender,  colourless,  not 
articulated,  straight  or  slightly 
undulated,  and  in  places  twisted 
in  irregular  spirals  with  pseudo- 
branchings.  The  development  can 
be  traced  from  the  cocci  to  rods  and 


FIG.  215.— BEGGIATOA  ALBA. 

A.  Threads  at  base  distinctly  linked,  partly  spiral.  B.  A  -thread,  spiral  in  its 
whole  length.  C,  D.  Fragments  detached  from  threads ;  immotile.  E.  Active 
spirillum-forms,  with  a  flagellum  at  either  end.  F,  G.  Thin  and  short  spiral 
forms.  H.  A  spiral  showing  the  individual  links,  x  540  (Zopf). 


whole  marshes  and  ponds  may  be 
coloured  blood-red  by  them. 

Spirillum  sanguineum,  rosaceum, 
violaceum,  monas  vinosa  and  Okenii, 
and  Rhabdomonas  rosea  are  pos- 
sibly phase-forms  of  Beggiatoa 
roseo-persicina. 


threads.  The  latter  are  at  the 
beginning  simple  threads,  which 
were  formerly  described  as  Lepto- 
thrix  parasitica,  or,  if  coloured  by 
impregnation  with  iron,  as  Lepto- 
thrix  ochracea.  Later  they  form 
false  branches  by  single  rods  turning 


544 


DESCRIPTION   OF   SPECIES. 


aside,  which  by  repeated  division 
lengthen  into  threads.  A  thread 
appears  to  be  first  composed  of 
long  rods,  then  of  short  rods,  and 
lastly  of  cocci.  The  iodine  reaction 
must  be  applied  to  distinguish  these 
forms,  especially  when  the  sheath 
of  the  threads  has  a  yellow,  rust- 
red,  olive-green,  or  dark  brown 
coloration.  The  cocci  may  grow 
into  rods  while  still  in  the  sheath, 
and  finally  become  leptothrix 
threads,  surrounded  by  a  delicate 
gelatinous  sheath,  from  which  the 


media  small  tufts,  about  1  to  3  /LI, 
and  floating  masses. 

Cladothrix  Fbrsteri  (ride  Strep- 
tothrix  Forsteri,  Cohn). 

Cladothrix  intricata  (Kussell). 
— Rods  and  filaments. 

Colonies  are  composed  of  a  net- 
work of  twisted  threads. 

Inoculated  in  the  depth  of  gela- 
tine fine  filaments  spread  out  from 
the  track  of  the  needle,  and  the 
gelatine  is  liquefied. 

Grown  on  agar  the  filaments 
penetrate  the  jelly. 


FIG.  216.— PHASE-FORMS  OF  BEGGIATOA  ROSEO-PERSICINA   (WARMING). 


false  branching  proceeds.  Frag- 
ments may  break  oft3,  which  are 
actively  motile,  and  appear  as 
vibrios,  spirilla,  and  spirochaeta- 
forms.  They  may  also  occur  in 
zooglcea  (Fig.  217). 

They  are  the  commonest  of  all 
bacteria  in  both  still  and  running 
water,  in  which  organic  substances 
are  present.  They  are  observed 
also  in  the  waste  water  of  certain 
manufactures,  such  as  sugar.  Arti- 
ficially they  can  be  cultivated  on 
infusions  of  rotting  algae  and  ani- 
mal substances,  forming  on  these 


In  broth  the  growth  is  abundant. 

They  were  isolated  from  sea 
dredgings. 

Cladothrix  invulnerabilis  (Ac- 
osta,  y  Grande  Rossi). — Filaments 
which  produce  in  gelatine  a  white 
thread,  and  liquefy  it  very  slowly. 

On  potato  the  growth  is  abundant 
and  chalky  in  appearance. 

In  milk  they  form  a  firm  yel- 
lowish pellicle  ;  and  in  broth  and  in 
water  the  growth  is  abundant. 

They  occur  in  water. 

Clostridium  butyricum  (vide 
Bacillus  bitty  ricus). 


DESCRIPTION   OF   SPECIES. 


545 


Clostridium  foetidum  (Libo- 
rius). — Rods  1  /j,  in  width,  singly  and 
in  filaments.  Spore-formation  re- 
sembles that  of  Bacillus  butyricus. 
They  are  anaerobic. 

Colonies  rapidly  liquefy  gelatine. 


gas-formation  with  unpleasant  smell 
and  splitting  up  of  the  jelly. 

They  were  isolated  from  earth. 

Crenothrix  Kiihniana  (Raben- 
horst). — Cocci,  rods,  and  thread- 
forms.  The  cocci  are  globular, 


FIG.  217.— CLADOTHRIX  DICHOTOMA. 

A.  Branching  schizomycete  :— (a)  Vibrio-form  ;  (6)  Spirillum-form  [slightly  mag- 

nified]. 

B.  A  screw-form  with  («)  Spirillum -form ;  (b)  Vibrio-form. 

C.  Long  spirochaeta-form. 

I).  Fragment  with  spirillum-form  at  one  end,  vibrio-form  at  the  other. 

E.  Screw-forms  : — (a)  continuous;  (b)  composed  of  rods;  (c)  composed  of  cocci. 

F.  Spirochseta-form  : — (a)  continuous;  (b)  composed  of  long  rods;  (c)  short  rods; 

(d)  cocci  (Zopf). 


On  agar  the  colonies  form  branch- 
ing processes  resembling  colonies  of 
Bacillus  cedematis  maligni. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  spreads  from  be- 
low upwards.  There  is  abundant 


1  to  6  n  in  diam.  The  threads  are 
colourless,  1*5  to  5  p.  thick,  and  club- 
shaped  at  the  extremity,  reaching 
a  diam.  of  6  to  9  p.  The  threads 
form  colonies  with  a  brick -red,  olive- 
green,or  dark-brown  to  brown- black 

35 


546 


DESCRIPTION   OF   SPECIES. 


coloration,  caused  by  impreg- 
nation with  oxide  of  iron.  The 
threads  are  distinctly  articulated, 
and  ensheathed.  The  segments  are 


set  free  when  the  sheath  bursts, 
and  develop  into  new  threads.  In 
other  cases  the  segments  remain 
enclosed,  and  subdivide  into  discs, 


FIG.  218.— CRENOTHRIX  KUHNIANA. 

«,  6,  c,  d,  e.  Cocci  in  various  stages  of  fission,  x  600. 

/.  Zooglcea  of  cocci,  x  600. 

(j.  Various  forms  of  zooglcea,  natural  size. 

h.  Colony  of  threads  composed  of  rods  grown  out  of  a  zooglcea  of  cocci. 

i — r.  Thread-forms  ;  some  straight,  others  spiral,  with  more  or  less  differentia- 
tion between  base  and  apex,  (r)  is  composed  of  short  rods  at  the  base,  and  above 
these  of  cylindrical  segments,  and  at  the  apex  these  segments  have  divided  into 
cocci,  x  600  (Zopf ). 


DESCRIPTION   OF   SPECIES. 


547 


which,  by  vertical  fission,  break  up 
into  globular  forms  (cocci).  These 
again  develop  into  new  threads, 
either  within  the  sheath,  eventually 
penetrating  it,  or  after  they  are  set 
free. 

The  micro-organism  appears  in 
little  whitish  or  brownish  tufts  in 
wells  and  drain-pipes,  and  it  not 
only  renders  drinking-water  foul, 
but  may  stop  up  the  narrower 
pipes. 

Diplococcus  albicans  ampins 
(Bumm). — Cocci  resembling  gono- 
cocci  but  much  larger,  singly  and 
in  tetrads. 

Colonies  are  prominent  and 
greyish-white. 

In  the  depth  of  gelatine  they 
produce  a  greyish-white  growth  in 
the  track  of  the  needle  and  on  the 
free  surface.  They  slowly  liquefy 
the  gelatine. 

They  were  obtained  from  the 
vaginal  mucous  membrane. 

Diplococcus  albicans  tardis- 
simus  (Bumm).  —  Cocci  morpho- 
logically identical  with  gonococci. 
They  grow  extraordinarily  slowly 
on  gelatine. 

They  form  very  minute  colonies, 
which  are  opaque  and  greenish- 
brown  in  colour. 

Inoculated  in  the  depth  of  gela- 
tine isolated  greyish- white  colonies 
develop  in  the  track  of  the  needle, 
and  on  the  free  surface  a  thin, 
white,  waxy  film  with  dentated 
edge. 

On  agar  the  growth  is  very 
similar. 

They  were  isolated  from  the 
vaginal  mucous  membrane. 

Diplococcus  citreus  conglome- 
ratus  (Bumm). — Cocci  in  pairs 
resembling  gonococci,  1  '5  \L  in  diam., 
in  tetrads  and  in  masses. 

Colonies  lemon-yellow ;  irregular 
in  form. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  develop  in  the  track 
of  the  needle,  and  liquefaction  com- 
mences at  its  upper  part. 

The  growth  on  the  free  surface 
is  yellow,  and  floats  on  the  liquefied 
gelatine  or  subsides  to  the  bottom 
of  the  liquefied  area. 


They  are  present  in  gonorrhoeal 
pus,  in  air  and  in  dust. 

Diplococcus  citreus  lique- 
faciens  (Unna).— Oval  cocci  -4  to 
•1  /i  in  diam.,  in  pairs,  tetrads, 
short  chains,  and  masses. 

Colonies  appear  in  the  form  of 
circular  discs,  at  first  greyish- white, 
later  lemon-yellow.  They  are  finely 
granular,  and  have  sharply  defined 
contours. 

Inoculated  in  the  depth  of  gela- 
tine, at  the  end  of  a  week  the 
growth  is  found  on  the  free 
surface,  forming  a  shining  yellow 
layer  ;  in  two  weeks  liquefaction 
commences,  and  the  growth  floats 
on  the  liquefied  gelatine  which  is 
also  yellowish  and  turbid. 

On  the  surface  of  agar  a  yellow- 
ish-brown layer  is  rapidly  formed. 

The  appearance  is  similar  on 
potato. 

They  were  isolated  in  cases  of 
eczema  seborrhosicum. 

Diplococcus  coryzae  (Hajek).— 
Large  diplococci. 

Colonies  are  white,  prominent. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  resembles  the 
pneumococcus. 

On  agar  a  white  layer  is  formed. 

They  are  probably  identical  with 
Friedliinder's  pneumococci. 

They  were  isolated  from  the 
mucus  in  acute  nasal  catarrh. 

Diplococcus  flavus  liquefaciens 
tardus. — Cocci  resembling  gono- 
cocci. 

Colonies  are  circular,  shining, 
and  chrome-yellow  in  colour. 

Inoculated  in  the  depth  of  gela- 
tine a  yellowish  growth  occurs 
along  the  needle  track,  and  also 
on  the  free  surface.  In  a  month 
the  surface  is  depressed,  but  the 
gelatine  is  not  liquefied  until  about 
two  months  have  elapsed. 

On  agar  a  yellowish-white  layer 
is  formed,  and  on  potato  the  colour 
is  more  pronounced. 

They  were  isolated  from  the 
skin  in  eczema  seborrhoeicum. 

Diplococcus  fluorescens  foeti- 
dus  (Klamann). — Cocci  in  pairs  and 
chains. 

Colonies     circular,     forming     a 


548 


DESCRIPTION    OF  SPECIES. 


brownish  deposit  surrounded  by 
liquefied  gelatine  which  has  a  violet 
or  greenish  tinge. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  along 
the  track  of  the  needle,  with  a 
hemispherical  excavation  of  the 
gelatine  at  the  upper  part.  An 
iridescent  film  floats  on  the  surface 
and  a  greenish  sediment  forms  at 
the  bottom  of  the  liquefied  area. 

On  agar  the  layer  is  brownish. 

On  potato  the  growth  is  granular, 
and  the  potato  in  the  vicinity  has 
a  bluish  colour. 

They  were  cultivated  from  the 
nasal  mucus. 

Diplocqccus  inter  cellularis 
meningitidis  (Weichselbaum).— 
Cocci  singly,  in  pairs,  tetrads  and 
masses.  They  grow  at  37°C. 

Colonies  on  agar  are  granular  and 
yellowish-brown. 

On  the  surface  of  agar  they  form 
a  greyish-white  viscid  growth.  In 
the  depth  of  agar  the  growth  only 
occurs  in  the  upper  part  of  the 
needle  track. 

On  blood  serum  and  broth  there 
is  very  little  growth,  and  none  on 
potato. 

Cultures  quickly  lose  their 
vitality. 

They  are  pathogenic  in  mice, 
guinea-pigs,  rabbits,  and  dogs. 

They  were  isolated  from  the  exu- 
dation in  cases  of  cerebro-spinal 
meningitis,  and  were  observed  in 
the  interior  of  pus  cells. 

Diplpcoccus  luteus  (Adametz). 
— Cocci  1'2  to  1*3  p  in  diam.,  singly 
and  in  chains.  Motile. 

Colonies  are  circular  and  slightly 
yellow,  and  granular.  Old  colonies 
are  bright  yellow. 

On  the  surface  of  gelatine  a 
growth  occurs  in  concentric  circles 
of  a  lemon-yellow  colour,  and  the 
gelatine  is  coloured  reddish-brown. 
After  several  weeks  liquefaction 
sets  in. 

On  agar  a  yellow  layer  forms,  and 
the  jelly  is  coloured  reddish-brown. 

On  potato  the  growth  changes 
from  yellow  to  brown.  Milk  is 
coagulated. 

They  were  obtained  from  water. 


Diplococcus  of  pneumonia  in 
horses  (Schutz). — Oval  cocci,  singly 
or  in  pairs,  capsulated. 

Colonies  small  and  white. 

In  the  depth  of  gelatine  a  row  of 
colonies  develops  in  the  track  of  the 
needle. 

On  agar  the  growth  is  composed 
of  transparent  droplets. 

Injection  into  the  lung  is  said  to 
produce  pneumonia,  ending  fatally 
in  eight  or  nine  days. 

They  are  pathogenic  in  rabbits, 
guinea-pigs,  and  mice. 

They  were  isolated  from  the 
lungs  of  a  horse  suffering  from 
pneumonia. 

Diplococcus  roseus  (Bumm). — 
Cocci  identical  in  description  with 
gonococci. 

Colonies  are  pink,  granular,  and 
irregular  in  form. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  grow  freely  in  the 
track  of  the  needle  and  on  the 
surface,  developing  a  pink  colour 
and  slowly  producing  liquefaction. 

They  are  present  in  the  air. 

Diplococcus  subflavus  (Bumm). 
— Diplococci  similar  to  gonococci. 

Colonies  greyish- white,  later 
yellow. 

They  grow  in  gelatine  and  on 
blood  serum,  and  liquefy  broth. 

They  produce  suppuration  when 
injected  subcutaneously  in  man. 

They  were  isolated  from  lochial 
discharges,  the  vesicles  of  pem- 
phigus, and  from  the  secretion  in 
colpitis  in  children. 

They  stain  by  Gram's  method. 

Hasmatococcus  bovis  (Babes).— 
Cocci  oval,  singly,  in  pairs,  and  in 
masses. 

Inoculated  in  the  depth  of  gela- 
tine minute  colonies  develop  in  the 
track  of  the  needle. 

On  agar  the  growth  is  composed 
of  transparent  droplets. 

On  potato  a  yellowish  shining 
film  is  formed. 

On  blood  serum  the  growth  is 
similar  to  that  on  agar. 

They  produce  a  fatal  result  in 
rabbits  and  guinea-pigs  in  a  week 
or  ten  days. 

They    were    isolated    from    the 


DESCRIPTION   OF   SPECIES. 


549 


blood  and  organs  of  cattle  which 
died  of  an  epidemic  disease  asso- 
ciated with  haemoglobinuria. 

Helicobacterium  aerogenes 
(Miller). — Bacilli  singly,  in  chains 
and  long  wavy  filaments.  Motile. 

Colonies  whitish,  varying  in  form. 

Inoculated  in  the  depth  of  gela- 
tine the  bacilli  give  rise  to  a  faintly 
yellow  growth  in  the  track  of  the 
needle,  and  an  almost  invisible, 
rapidly  growing  layer  on  the 
surface. 

On  potato  the  growth  is  dry  and 
brownish. 

They  were  isolated  from  the 
healthy  intestinal  tract. 

Leptothrix  buccalis  (Robin). — 
Long,  thin  threads,  -7  to  1  /x  broad, 
colourless,  often  united  in  thick 
bundles  or  felted  together.  Masses 
of  cocci  occur  with  the  threads, 
and  the  threads  themselves  are  com- 
posed of  long  rods,  short  rods,  and 
cocci.  The  threads  may  break  up 
into  spiral,  vibrio,  and  spirochaeta 
forms.  The  last-named  occur  in 
large  numbers  in  the  mouth,  and 
have  been  named  Spirochceta  buc- 
calis. Leptothrix  buccalis  is  found 
in  teeth  slime,  and  is  believed 
to  be  intimately  connected  with 
dental  caries.  The  threads  pene- 
trate the  tissue  of  the  teeth,  after 
the  enamel  has  been  acted  upon  by 
acids  generated  by  the  fermentation 
of  food.  The  short  rods,  long  rods, 
cocci,  leptothrix-forms,  and  screw- 
forms  are  found  in  the  dental 
canals. 

The  threads  of  Leptothrix  buc- 
calis have  a  special  staining  reaction 
(Leber).  They  become  coloured  if 
placed  in  an  acid  medium  with 
iodine  ;  if  the  medium  be  alkaline, 
it  must  first  be  acidified  with  very 
dilute  hydrochloric  acid  or  acetic 
acid.  The  contents  are  stained 
violet,  and  contrast  with  the  sheath 
and  septa,  which  remain  uncoloured. 

Leptothrix  buccalis  (Vignal).-— 
Rods  1  to  1-5  \i  in  width,  1-6  to 
30  /u.  in  length. 

Colonies  greyish-white,  promi- 
nent and  furrowed. 

Inoculated  in  the  depth  of  gela- 
tine a  filament  forms  in  the  track 


of  the  needle,  and  a  growth  occurs 
on  the  free  surface.  Liquefaction 
sets  in  at  the  upper  part,  forming 
a  cup-shaped  cavity,  and  a  bluish 
skin  floats  upon  the  liquid.  The 
liquefaction  gradually  extends  to 
the  side  of  the  tube,  and  a  deposit 
is  formed  at  the  bottom  of  the 
liquefied  gelatine. 

On  agar  the  layer  is  white, 
wrinkled  and  transparent,  and  later 
yellowish. 

In  broth  there  is  turbidity,  but 
no  skin  on  the  surface. 

On  potato  the  growth  is  greyish- 
white. 

They  are  occasionally  present 
in  the  mouth  in  health,  and 
are  possibly  identical  with  lepto- 
thrix  buccalis  (Robin). 

Leptothrix  gigantea  (Miller). — 
Long  rods,  short  rods  and  cocci 
can  be  observed  in  the  same  thread. 
There  are  also  screw-threads,  which 
may  take  the  form  of  spirals, 
vibrios,  or  spirochaetae.  The  threads 
increase  in  diameter  from  base 
to  apex  ;  corresponding  with  the 
thickness  of  the  threads,  the  rods 
and  cocci  show  different  dimensions. 

They  have  been  observed  in  the 
diseased  teeth  of  dogs,  sheep,  cats 
and  other  animals. 

Leuconostoc  mesenteroides, 
Cienkowski  (Gomme  de  siicrerie, 
Froschlaichpilz,  Frogspawn  fungus). 
— Cocci  and  rods  singly,  in  chains, 
and  in  zoogloea,  surrounded  by  a 
thick  gelatinous  envelope.  The  life- 
history  has  been  very  thoroughly 
investigated.  The  spores,  1*8  to 
2  p.  in  diam.,  are  of  a  round  or 
ellipsoidal  form,  with  thick  mem- 
brane and  shining  contents.  The 
outer  membrane-layer  bursts,  and  a 
middle  lamella  oozes  out,  and  forms 
a  thick  gelatinous  envelope,  while 
the  inner  layer  remains  adherent 
to  the  plasma.  Thus  the  spore- 
germination  leads  to  the  formation 
of  a  coccus  with  a  gelatinous  en- 
velope. The  coccus  then  elongates 
into  a  short  rod-form,  and  the 
gelatinous  envelope  becomes  ellip- 
soidal. The  rod  divides  into  two 
cocci,  and  each  of  these  lengthens 
into  a  rod  and  divides.  By  repetition 


550 


DESCRIPTION   OF  SPECIES. 


of  this  process  a  chain  of  cocci 
results,  encased  in  a  cylindrical 
or  ellipsoidal  envelope.  The  chains 
increase  in  length,  become  twisted 
up,  and  eventually  fall  apart  into 
pieces  of  various  lengths. 

In  nourishing  liquids  a  great 
number  of  little  masses  are  formed, 
which  adhere  together,  and  produce 
pseudo-parenchymatous  structures. 
These  latter  may  join  together, 
forming  still  larger  agglomerations. 


This  micro-organism  occurs  occa- 
sionally in  beet-root  juice  and  the 
molasses  of  sugar-makers,  forming 
large  gelatinous  masses  resembling 
frog-spawn.  The  vegetation  is  so 
rapid  that  forty-nine  hectolitres  of 
molasses,  containing  10  per  cent, 
of  sugar,  were  converted  within 
twelve  hours  into  a  gelatinous 
mass  ;  consequently,  it  is  a  for- 
midable enemy  of  the  sugar  manu- 
facturers. 


FIG.  219.— LEUCONOSTOC  MESENTEROIDES. 

1.  Spores.  2.  Spores  after  germination,  showing  gelatinous  envelope.  3,  4,  5,  6. 
Increase  by  division.  7.  Glomerular  form  of  zooglcea.  8.  Section  of  an  old 
kowskif  Z°°gloea"  9*  Cocci  chains  with  arthrospores  (Tieghem  and  Cien- 


The  masses  of  zooglcea  are  of 
almost  a  cartilaginous  consistency, 
and  admit  of  sections  being  made 
with  a  razor.  After  a  long  time 
the  envelope  liquefies,  and  the  cocci 
are  set  free  ;  the  latter  introduced 
into  fresh  nourishing  media  develop 
new  colonies.  In  the  chains  some 
of  the  cocci  become  enlarged  with- 
out changing  their  form.  These 
acquire  the  properties  of  spores, 
and  are  firthrospores. 


Micrococcus    acidi   lactici 

(Marpmann). — Large  cocci,  singly 
and  in  pairs. 

Colonies  yellowish-white. 

On  the  surface  of  gelatine  the 
cocci  produce  a  yellow  layer. 

They  grow  in  milk,  producing  a 
reddish  colour,  and  coagulation  due 
to  the  formation  of  lactic  acid. 

They  were  isolated  from  milk. 

Micrococcus  acidi  lactici  lique- 
faciens  (Kreuger).— Cocci  oval,  1 


DESCRIPTION   OF  SPECIES. 


551 


to  1*5  p.  in  diam.,  in  pairs  and  in 
tetrads. 

Colonies  white. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  produce  a  granular 
filament,  changing  in  two  or  three 
days  to  liquefaction  in  the  form  of 
a  funnel ;  later,  a  wrinkled  mem- 
brane floats  on  the  surface  of  the 
liquid. 

In  milk  they  produce  lactic  acid. 

They  were  isolated  from  butter 
which  had  turned  cheesy. 

Micrococcus  aerogenes  (Miller). 
— Oval  cocci. 

Colonies  are  dark  and  regular  in 
contour,  but  have  a  peculiar  spotted 
appearance. 

Inoculated  in  the  depth  of  gela- 
tine a  brownish-yellow  growth 
occurs  along  the  track  of  the  needle, 
and  on  the  free  surface  a  white 
button-like  elevation.  After  a  time 
the  gelatine  is  slowly  liquefied. 

On  agar  a  yellowish- white  pulpy 
layer  forms,  and  a  similar  growth 
appears  on  potato. 

They  resist  the  action  of  acids,  so 
that  the  presence  of  gastric  juice 
does  not  impede  their  develop- 
ment. 

They  were  obtained  from  the 
intestine. 

Micrococcus  agilis  (Ali-Cohen). 
—Cocci  1  /x  in  diam.,  singly,  in 
pairs,  tetrads,  and  in  chains.  They 
are  motile,  and  possess  flagella. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  in  the  track  of  the 
needle,  and  produce,  after  two  or 
three  weeks,  liquefaction  or  excava- 
tion of  the  jelly. 

On  agar  and  potato  the  growth 
is  pink. 

They  occur  in  water. 

Micrococcus  agilis  citreus 
(Menge). — Cocci  in  pairs,  chains 
and  masses.  They  are  motile,  and 
each  coccus  possesses  a  single  fla- 
gellum. 

Colonies  appear  surrounded  by 
clouded  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  scanty  growth  in  the 
track  of  the  needle,  and  on  the  sur- 
face a  bright  yellow  patch. 

On  agar  they  form  a  yellow  layer, 


which  is  viscid,  and  may  be  drawn 
out  in  long  threads. 

In  broth  they  produce  cloudiness 
and  a  viscous  deposit. 

The  growth  on  potato  is  bright 
yellow. 

Milk  is  not  coagulated. 

They  were  isolated  from  an  in- 
fusion of  peas. 

Micrococcus  albus  liquefaciens 
(Besser). — Large  cocci  in  chains  and 
in  masses.  They  are  anaerobic. 

Colonies  on  agar  exhibit  concen- 
tric rings  of  different  shades  of 
brown. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction  in 
the  track  of  the  needle. 

They  occur  in  mucus  from  the 
nose. 

Micrococcus  amylivorus  (Bur- 
rill). — Oval  cocci,  1  to  1'4  p.  long, 
•7  n  broad,  singly,  in  pairs,  and  rarely 
in  fours,  never  in  chains,  are  found 
embedded  in  an  abundant  mucilage 
which  is  very  soluble  in  water. 

They  have  been  described  as  pro- 
ducing the  so-called  "  fire  blight " 
of  the  pear  tree  and  other  plants. 

Micrococcus  aquatilis  (Bolton). 
— Small  cocci  in  masses. 

Colonies  circular,  prominent,  and 
pure-white. 

Inoculated  in  the  depth  of  gela- 
tine, there  is  a  white  growth  in  the 
track  of  the  needle  and  also  on  the 
free  surface. 

On  agar  the  growth  is  white. 

They  occur  in  water. 

Micrococcus  aquatilis  invisi- 
bilis  (Vaughan). — Cocci  oval. 

Colonies  brown. 

In  gelatine  there  is  a  slight 
growth  in  the  track  of  the  needle, 
and  a  more  abundant  growth  on 
the  free  surface. 

On  agar  they  form  a  white  film. 

On  potato  the  growth  is  in- 
visible. 

They  occur  in  water. 

Micrococcus  aurantiacus 
(Cohn). — Cocci  spherical  or  oval, 
1-3  to  1-5  p.  in  diam.,  singly,  in  pairs, 
and  in  groups. 

Colonies  orange-yellow. 

Inoculated  in  gelatine  they  form 
minute  colonies  in  the  track  of  the 


552 


DESCRIPTION   OF  SPECIES. 


needle,  and  a  prominent  hemi- 
spherical yellow  growth  on  the  free 
surface. 

On  agar  the  growth  is  orange- 
yellow,  and  on  potato  yellow  and 
slimy. 

They  occur  in  water. 

Micrococcus  botryogenus 
(Johne,  Rabe). — Cocci  1  to  1'5  p,  in 
diam.,  in  wavy  chains. 

Colonies  circular,  sharply  defined. 
At  first  silver-grey,  later  yellowish- 
grey  with  metallic  lustre,  they 
produce  an  odour  like  that  of 
strawberries. 

Inoculated  in  the  depth  of  gela- 
tine a  greyish- white  filament  de- 
velops, with  slight  liquefaction  of 
the  gelatine  ;  later,  it  becomes 
milk-white,  and  at  its  upper  part  a 
characteristic  bubble  appears. 

They  make  hardly  any  growth 
on  agar. 

On  potato  they  grow  very  abun- 
.dantly,  forming  a  yellowish  layer 
with  the  same  odour  as  the  colonies 
on  plate  cultivations. 

Inoculated  guinea-pigs  die  of 
septicaemia  ;  in  sheep  and  goats 
severe  inflammation  spreads  from 
the  point  of  inoculation.  Mice  are 
immune.  In  horses  an  inflamma- 
tory oedema  is  at  first  set  up, 
followed  in  four  to  six  weeks  by 
the  formation  of  new  growths, 
which  sometimes  suppurate  and 
contain  large  numbers  of  micro- 
cocci. 

They  were  found  in  tumours  of 
the  spermatic  cord  and  of  the 
connective  tissue  in  other  parts  in 
horses. 

Micrococcus  candicans 
(Fliigge). — Cocci  which  collect  in 
masses. 

In  plate-cultivations  they  form 
in  two  or  three  days  milk-white 
colonies ;  while  those  below  the 
surface  of  the  gelatine  are  yellow- 
ish. Under  a  low  power  the  deep 
colonies  are  quite  circular,  with 
smooth  margins,  of  a  blackish-brown 
colour,  and  very  slightly  granular  ; 
the  superficial  colonies  are  quite 
irregular  in  outline,  and  are  finely 
granular. 

Cultivated  in  test-tubes  they  form 


a  white  nail-shaped  cultivation. 
They  were  isolated  from  contami- 
nated plate-cultivations. 

They  occur  in  the  air. 

Micrococcus  candidus(Cohn).— 
Cocci  forming  snow-white  points 
and  spots  upon  slices  of  cooked 
potato. 

Micrococcus  carneus  (Zimmer- 
mann). — Cocci  *8  /z  in  diam.,  occur- 
ring in  masses. 

Colonies  circular,  greyish-white, 
with  the  centre  tinged  with  red. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  white,  granular 
filament  in  the  track  of  the  needle, 
and  a  pale  pink  layer  on  the  free 
surface. 

On  the  surface  of  oblique  gela- 
tine a  flesh-coloured  layer  develops, 
which  later  assumes  a  violet  colour. 

On  agar  the  growth  is  similar. 

On  potato  the  growth  is  abundant 
and  red  in  colour. 

They  were  isolated  from  water. 

Micrococcus  cerasinus  siccus 
(List). — Cocci  *25  to  '12  p  in  diam., 
singly  and  in  pairs.  They  can  best 
be  cultivated  at  37°  C. 

On  agar  they  form  a  cherry-red 
layer,  and  a  similar  growth  on 
potato. 

The  pigment  is  insoluble  in  alco- 
hol, ether,  and  water,  and  is  not 
destroyed  by  acids  or  alkalies. 

They  occur  in  water. 

Micrococcus  cereus  albus 
(p.  178). 

Micrococcus  cereus  flavus 
(p.  178). 

Micrococcus  cinnabareus 
(Fliigge). — Large  cocci  occurring  in 
twos,  threes,  and  fours. 

Colonies  develop  very  slowly,  and 
are  punctiform,  and  bright  red  at 
first,  and  afterwards  reddish-brown. 

The  cocci  inoculated  on  the  sur- 
face of  gelatine  form  a  heaped-up, 
red-coloured  growth. 

They  were  found  contaminating 
old  cultivations. 

Micrococcus  citreus  (List).— 
Cocci  1'5  to  2-2  /j.  in  diam.,  singly, 
in  pairs  and  chains. 

Colonies  are  irregular  in  form, 
moist  and  shining,  and  yellowish  in 
colour. 


DESCRIPTION    OF   SPECIES. 


553 


In  the  depth  of  gelatine  the 
growth  is  very  scanty. 

On  the  surface  of  agar  the  growth 
is  yellowish. 

On  potato  the  growth  is  similar 
but  more  abundant. 

Micrococcus  concentricus 
(Zimmermann). — Cocci  -9/zin  diam., 
in  masses. 

Colonies  bluish-grey. 

Inoculated  in  the  depth  of  gela- 
tine there  is  no  growth  in  the  track 
of  the  needle,  but  concentric  rings 
form  on  the  free  surface. 

On  agar  the  growth  is  greyish- 
white  and  smooth. 

On  potato  yellowish  and  slimy. 

They  occur  in  water. 

Micrococcus  cremoides  (Zim- 
mermann).— -Cocci  '8  /z  in  diam., 
occurring  in  masses. 

Colonies  are  spherical,  granular 
and  yellowish.  The  margins  are 
dentated  or  irregular,  and  processes 
extend  into  the  surround  ing  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the 
track  of  the  needle  in  a  few  days. 
A  yellow  growth  floats  on  the 
liquefied  gelatine,  and  a  yellowish 
mass  subsides  to  the  bottom  of  the 
liquid. 

On  agar  a  smooth  shining  layer 
is  formed,  and  on  potato  the  growth 
is  abundant. 

They  occur  in  water. 

Micrococcus  crepusculum 
(Cohn.  Monas  crepusculum,  Ehren- 
berg.  Jfikrokokken  in  faulenden 
Substraten,  Fliigge). — Round  or 
short  oval  cells,  scarcely  2  //,  in 
diam.  ;  singly  or  in  zoogloea. 

They  occur  in  various  infusions 
and  putrefying  fluids  in  company 
with  Bacterium  termo. 

Micrococcus  cumulatus  tenuis 
(Besser). — Large  cocci,  oval  ;  in 


Colonies  on  agar  have  a  brown 
nucleus. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  white  filament, 
and  on  the  surface  a  transparent 
layer. 

In  broth  there  is  an  abundant 
deposit,  and  the  supernatant  liquid 
is  clear. 


They  occur  in  mucus  from  the 
nose. 

Micrococcus  endocarditidis  ru- 
gatus  (Weichselbaum). — Cocci  re- 
sembling pyogenic  staphylococci. 

Colonies  have  a  brown  or  yellow- 
ish-brown nucleus. 

In  the  depth  of  agar  there  is  a 
slight  growth  in  the  track  of  the 
needle,  and  a  wrinkled,  waxy  layer 
on  the  surface. 

On  potato  the  growth  is  dry  and 
brownish. 

On  blood  serum  the  growth  is 
colourless  and  adherent. 

In  jectedsubcutaneously,  they  pro- 
duce, in  rabbits,  local  swelling  and 
redness,  and  suppuration  in  guinea- 
pigs.  Injected  into  the  veins  after 
injury  to  the  aortic  valves,  they 
produce  endocarditis. 

They  were  isolated  from  a  case 
of  ulcerative  endocarditis. 

Micrococcus  fervidosus  (Ada- 
metz). — Cocci  '6  /A  in  diam.,  in  pairs 
and  in  masses. 

The  deep  colonies  are  pale-yellow, 
and  look  like  watery  droplets  ;  but 
superficial  colonies  are  granular  and 
irregular  with  jagged  edges. 

Inoculated  in  the  depth  of  gela- 
tine a  granular  filament  develops  in 
the  track  of  the  needle,  and  on  the 
surface  a  circular  patch  with  den- 
tated margin. 

On  agar  the  growth  is  white  and 
slimy,  and  on  potato  greyish - 
white. 

They  occur  in  water. 

Micrococcus  Finlayensis 
(Sternberg). — Cocci  '5  to  '1  p.  in 
diam.,  singly,  in  pairs,  tetrads,  and 
in  masses. 

In  the  depth  of  gelatine  they 
produce  a  growth  in  the  track  of 
the  needle,  with  liquefaction  at 
the  upper  part  with  a  pale-yellow 
deposit. 

On  agar  the  growth  is  pale-yellow. 

They  were  isolated  from  the  liver 
in  a  fatal  case  of  yellow  fever. 

Micrococcus  flavus  desidens 
(Fliigge). — Cocci  singly,  in  pairs, 
or  chains  of  a  few  elements. 

Colonies  yellowish-white. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  along  the  track  of 


554 


DESCRIPTION   OF  SPECIES. 


the  needle,  and  form  a  yellowish- 
brown  layer  at  the  point  of  punc- 
ture. 

Later  liquefaction  sets  in,  and  a 
deposit  forms  at  the  bottom  of  the 
turbid  liquid. 

They  occur  in  air  and  in  water. 

Micrococcus  flavus  lique- 
faciens  (Fliigge).— Cocci  mostly  in 
twos  and  threes,  also  in  masses. 

Small  yellow  colonies  appear  after 
two  or  three  days,  which  have  a 
shallow  depressed  zone  surrounding  j 
them.  Under  a  low  power  they 
are  granular  and  yellowish-brown, 
with  lines  radiating  from  the  centre 
to  the  circumference.  Later  they 
liquefy  the  gelatine,  and  coalesce. 

Inoculated  in  the  depth  of  gelatine 
the  cocci  produce  spherical  yellow 
colonies  in  two  days  along  the  track 
of  the  needle.  These  become  con- 
fluent, and  at  the  end  of  eight  days 
the  whole  of  the  jelly  has  become 
liquid  ;  later  the  upper  part  becomes 
clear,  and  a  yellow  mass  subsides  to 
the  bottom  of  the  tube. 

They  occur  in  air  and  in  water. 

Micrococcus  flavus  tardigra- 
dus  (Fliigge). — Large  cocci  showing 
at  times  peculiar  dark  poles  ;  gener- 
ally arranged  in  masses. 

Colonies  develop  slowly ;  the 
superficial  ones  have  a  smooth  wax- 
like  surface  with  projecting  centre  ; 
those  below  the  surface  are  of  a 
dark  chrome-yellow  colour,  and  are 
round  or  oval. 

Inoculated  in  gelatine  the  cocci 
develop  slowly  along  the  track  of 
the  needle,  forming  small  isolated 
colonies ;  the  gelatine  is  not 
liquefied. 

They  occur  in  air  and  in  water. 

Micrococcus  fcetidus(Klamann).  , 
— Cocci  singly,  in  pairs,  and  short  j 
chains  and  masses. 

Colonies  circular  or  oval,  white. 

Inoculated  in  the  depth  of  gela-   | 
tine  a  pure  white,  shining  growth  j 
forms  in  concentric  circles  at  the  ! 
point  of  puncture,  and  develops  a  [ 
brownish  colour  ;  and  liquefaction 
occurs  after  a   time,  and  extends 
along  the  needle  track. 

A  white  layer  spreads  over  the 
surface  of  agar. 


On  potato  the  growth  is  slimy 
and  grey  in  colour,  with  a  red  tinge. 

Cultures  produce  an  odour  like 
that  of  ozaena. 

They  were  isolated  from  the 
nose. 

Micrococcus  ftetidus  (Rosen- 
bach). — Small  oval  cocci. 

Cultivated  in  agar-agar  they 
develop  gas-bubbles  and  a  foetid 
odour.  They  were  isolated  from 
carious  teeth. 

Micrococcus  Freudenreichi 
(Guillebeau). — Large  cocci,  singly 
and  in  chains. 

Colonies  are  granular  and  puncti- 
form. 

In  broth  turbidity  is  produced, 
and,  later,  a  flocculent  deposit. 

On  potato  a  shining  film  develops, 
yellowish  or  brownish-yellow  in 
colour. 

In  milk  the  cultures  become 
viscous,  and  can  be  drawn  out  into 
threads  several  yards  in  length. 

They  were  isolated  from  milk 
with  viscous  fermentation. 

Micrococcus  fuscus  (Maschek). 
— Cocci  oval. 

Colonies  pale-brown  or  black. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  slight  growth  along 
the  track  of  the  needle,  and  a  brown 
layer  forms  on  the  surface  followed 
by  liquefaction. 

On  potato  the  growth  is  brown 
or  brownish-black  and  slimy. 

Cultures  give  off  an  odour  of 
putrefaction. 

They  occur  in  water. 

Micrococcus  gingiyae  pyogenes 
(Miller). — Large  cocci,  singly  and 
in  pairs. 

Colonies  spherical,  with  sharp 
contours. 

Inoculated  in  the  depth  of  gela- 
tine there  is  an  abundant  growth 
along  the  track  of  the  needle  and 
on  the  free  surface. 

On  agar  a  thick  film  develops, 
with  a  faint  tinge  of  purple  by 
transmitted  light. 

Injected  into  mice  subcutaneously 
they  produce  local  suppuration,  and 
sometimes  death.  Injected  into 
the  peritoneal  cavity  they  produce 
peritonitis  and  death. 


DESCRIPTION   OF   SPECIES. 


555 


They  were  isolated  from  an 
abscess  of  the  gums. 

Micrococcus  gonorrhoeas 
(p.  190). 

Micrococcus  havaniensis 
(Sternberg). — Cocci  4'5  \i  in  diam. 

The  colonies  are  circular  and  of 
a  blood-red  colour. 

The  cocci  inoculated  in  the  depth 
of  gelatine  produce  a  colourless 
growth  in  the  track  of  the  needle 
and  a  carmine  patch  on  the  surface. 

On  agar  and  on  potato  they  form 
a  thick  irregular  carmine  layer. 

Micrococcus  in  Biskra-button 
(Heydenreich). — Cocci  in  pairs, 
•86  to  1  p.  in  length,  occasionally 
tetrads  :  capsulated. 

Inoculated  in  the  depth  of  gela- 
tine they  form  a  greyish-white  fila- 
ment composed  of  closely  packed 
colonies,  and  a  yellowish- white  film 
on  the  free  surface.  Liquefaction 
commences  at  the  upper  part  of  the 
needle  track  in  a  few  days,  forming 
a  funnel  which  extends  until,  in 
two  weeks,  the  gelatine  is  com- 
pletely liquefied. 

On  the  surface  of  agar  a  shining 
white  or  yellowish-white  layer  de- 
velops in  twenty-four  hours. 

On  potato  the  growth  is  similar. 

Inoculations  are  said  to  produce 
in  rabbits,  dogs,  fowls,  sheep  and 
horses  a  morbid  condition  of  the 
skin  similar  to  the  disease  known  as 
Biskra-button  or  Pendjeh  sore.  In 
man  they  produce  suppuration  when 
rubbed  on  the  skin. 

They  were  isolated  from  the 
disease  known  as  Pendjeh  sore, 
Biskra-button  or  clou  de  Btskra. 

Micrococcus  in  gangrenous 
mastitis  in  sheep.— Cocci  singly, 
in  pairs,  and  in  masses. 

Colonies  are  spherical,  white,  and 
under  a  low  power  have  a  brown 
nucleus  and  transparent  margin. 

The  cocci  inoculated  in  the  depth 
of  gelatine  produce  a  conical  area 
of  liquefied  jelly  with  a  copious 
white  deposit. 

On  agar  they  produce  a  white 
layer,  which  later  turns  yellowish 
in  colour. 

On  potato  they  form  a  greyish 
growth. 


Injected  into  the  mamary  gland 
of  sheep  they  produce  inflammatory 
oedema,  and  a  fatal  result  in 
twenty-four  to  forty-eight  hours. 

In  rabbits  they  are  pyogenic. 

They  were  isolated  from  the  milk 
in  cases  of  gangrenous  mastitis  in 
sheep. 

Micrococcus  in  infectious 
p  1  e  u  r  o-pneumonia  (Poels  and 
Nolen)— p.  242. 

Micrococcus  in  influenza  (Fis- 
chel). — Cocci  from  1  to  1-25  p  in 
diam.,  singly,  in  pairs,  and  chains. 

Extremely  minute  colonies  appear 
in  three  days. 

Inoculated  in  the  depth  of  gela- 
tine a  milk-white  filament  forms 
along  the  track  of  the  needle.  Lique- 
faction commences  in  four  days 
at  the  upper  part,  and  extends 
slowly. 

On  agar  the  colonies  are  pure- 
white. 

On  potato  the  growth  is  yellowish- 
white. 

They  do  not  grow  on  blood  serum 
or  in  milk. 

Intravenous  injection  in  dogs  is 
said  to  produce  symptoms  like 
distemper. 

They  were  obtained  from  the 
blood  in  cases  of  influenza. 

Micrococcus  in  influenza 
(Kircbner). — Cocci  in  pairs  and 
chains  ;  capsulated.  They  grow  at 
37°  C. 

The  colonies  are  transparent, 
whitish. 

On  the  surface  of  agar  there  is 
an  abundant  growth,  but  it  is 
limited  in  the  depth  of  the  jelly. 

Inoculation  experiments  were 
inconclusive. 

They  were  obtained  from  the 
sputum  in  cases  of  influenza. 

Micrococcus  in  pemphigus 
(Almquist). — Cocci  '5  to  1  /i  in 
diam.,  singly  and  in  pairs  ;  identi- 
cal with  Staphylococcus  pyogenes 
aureus. 

•  The  cocci  vaccinated  on  the  arm 
are  said  to  have  produced  bullae. 

They  were  isolated  from  pem- 
phigoid  bullas  in  children. 

Micrococcus  in  pemphigus 
(Demme). — Cocci  -8  to  1*4  p.  in 


556 


DESCRIPTION   OF   SPECIES. 


diam.,  singly,  in  pairs,  and  in 
masses.  They  can  be  cultivated 
at  37°  C. 

The  colonies  on  agar  are  milk- 
white  and  prominent.  Later,  off- 
shoots occur  from  the  margin, 
producing  a  resetted  appearance. 

Inoculated  in  the  depth  of 
gelatine  the  cocci  form  clubbed  or 
stalactitic  out -growths  from  the 
filament  which  develops  in  the 
track  of  the  needle. 

On  the  surface  of  agar  a  creamy 
layer  is  formed  with  similar  off- 
shoots. 

Injected  into  the  lungs  of  guinea- 
pigs  they  are  said  to  produce 
broncho-pneumonia. 

They  were  obtained  from  the 
bullae  in  acute  pemphigus. 

Micrococcus  in  pneumonia 
(Manfredi). — Oval  cocci  -6  to  1  /* 
in  width,  1  to  1'5  p  in  length,  singly, 
in  pairs,  and  short  chains. 

Colonies  on  gelatine  are  circular, 
whitish,  and  later  spread  out  and 
become  bluish  by  transmitted  light, 
and  of  a  pearly  lustre  by  reflected 
light. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  limited  growth  along 
the  track  of  the  needle. 

On  blood  serum  they  form  a 
shining,  granular,  faintly  greenish- 
yellow  layer. 

They  also  can  be  cultivated  on 
potato  and  in  broth. 

They  are  pathogenic  in  dogs, 
rabbits,  guinea-pigs,  mice  and  birds. 
Birds  die  in  a  few  days  ;  mammals 
in  from  one  to  three  weeks.  After 
death  new  growths  composed  of 
granulation  tissue  are  found  in  the 
internal  organs,  varying  in  size 
from  a  millet  seed  to  a  pea.  They 
were  obtained  from  the  sputum  of 
pneumonia  complicating  measles. 

Micrococcus  in  progressive 
abscess  formation  in  rabbits 
(Koch).— Cocci  only  about  '15  p.  in 
diam.,  principally  in  thick  zoogloea. 
The  disease  was  induced  by  the  in- 
jection into  rabbits  of  decomposing 
blood.  At  the  place  of  injection  a 
spreading  abscess  formed,  which  was 
fatal  to  the  animal  in  about  twelve 
days.  No  bacteria  were  observed 


:  in  the  blood,  but  in  the  walls  of  the 
abscess  thick  masses  of  cocci  were 
found.  The  pus  is  infectious, 
causing  the  same  disease  in  healthy 
rabbits. 

Micrococcus  in  pyaemia  in 
rabbits  (Koch). — Round  cocci  and 
diplococci  "25  /u,  in  diam. 

The  disease  was  produced  by  the 
subcutaneous  injection,  in  a  rabbit, 
of  distilled  water  in  which  the  skin 
of  a  mouse  had  been  macerated. 
At  the  autopsy  there  were  found 
great  infiltration  around  the  site  of 
injection,  peritonitis^  and  accumu- 
lations in  the  liver  and  lungs  ;  in 
short,  the  appearances  of  pyaemia. 


FIG.  220.— MICKOCOCCUS  IN  PYAEMIA 
IN  RABBITS  :  VESSEL  FROM  THE 
CORTEX  OF  THE  KIDNEY  x  700. 
a,  Nuclei  of  the  vascular  wall ; 
c,  i  Masses  of  micrococci  adherent 
to  the  wall  and  enclosing  blood - 
corpuscles  (Koch). 

In  the  capillaries  of  the  organs 
examined,  masses  of  cocci  were 
observed  enclosing  blood-corpuscles. 
Fresh  inoculations  in  rabbits  with 
exudation-fluid,  or  blood  from  the 
heart,  reproduced  the  same  disease. 
Micrococcus  in  septicaemia  in 
rabbits. — Ellipsoidal  cocci  -8  to  1 

j  IL  in  largest  diam.  The  disease 
was  produced  by  the  injection  of 

j    putrid  meat  infusion.     After  death 

'  slight  oedema  was  noted  at  the  site 
of  injection,  slight  extravasation  of 
blood,  and  great  enlargement  of 
the  spleen.  No  emboli  or  peri- 
tonitis resulted.  Masses  of  cocci 

1    were   found   in   the    capillaries   of 


DESCRIPTION   OF   SPECIES. 


557 


different  organs,  especially  in  the 
glomeruli  of  the  kidneys.  Rabbits 
and  mice  inoculated  with  blood 
from  the  heart  proved  susceptible 
to  the  disease. 

Micrococcus  in  syphilis  (Disse 
and  Taguchi). — Cocci  and  diplo- 
cocci. 

They  produce  a  greyish-white 
growth  in  nutrient  media. 

They  are  said  to  produce  inflam- 
matory changes  in  the  internal 
organs  and  disease  of  the  blood- 
vessels when  inoculated  in  dogs, 
rabbits  and  sheep. 

They  were  obtained  from  the 
blood  in  cases  of  syphilis. 

Micrococcus  in  trachoma 
(p.  190). 

Micrococcus  in  yellow  fever 
(p.  260). 

Micrococcus  lactis  viscosus 
(Conn). — Cocci  in  pairs  and  chains. 

Colonies  circular  and  granular. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  begins  at  the  upper 
part  of  the  needle  track,  and  extends 
until  the  gelatine  is  completely 
liquefied.  The  liquefied  gelatine  is 
viscous,  and  may  be  drawn  out  in 
long  threads. 

On  the  surface  of  agar  they  form 
a  white,  shining  layer. 

In  broth  there  is  an  abundant 
growth,  and  a  film  on  the  surface. 

They  coagulate  milk,  producing 
butyric  acid,  and  giving  it  a  bitter 
taste.  They  were  obtained  from 
bitter  cream. 

Micrococcus  luteus  (Cohn).— 
Oval  cocci  1  to  1'2  /*  in  diam. 

Colonies  yellow,  with  irregular 
contours,  and  granular. 

Inoculated  in  the  depth  of  gela- 
tine a  granular  filament  develops  in 
the  track  of  the  needle;  and  on  the 
free  surface  a  yellow  patch. 

On  agar  the  growth  is  slimy  and 
yellow. 

On  potato  the  growth  is  yellow, 
and  after  a  time  wrinkled. 

The  pigment  is  insoluble  in  water, 
ether  and  alcohol,  and  not  destroyed 
by  acids  or  alkalies. 

They  occur  in  water. 

Micrococcus  luteus  (Schrdter). 
— Cocci  similar  in  size  to  the  above, 


elliptical,  with  highly  refractive  cell 
contents. 

They  form  yellow  drops  of  1  to 
3  mm.  diam.  on  boiled  potato  ;  and 
a  thick,  wrinkled,  yellow  skin  on 
nutrient  liquids. 

The  colouring-matter  is  insoluble 
in  water,  and  unchanged  by  sul- 
phuric acid  or  alkalies. 

Micrococcus  ochroleucus 
(Prove). — Cocci  *5  to  '8  /M  in  diam., 
singly,  in  pairs,  and  short  chains. 

Colonies  minute  and  colourless, 
with  crenated  margin,  from  which, 
later,  processes  extend  into  the  gela- 
tine, while  the  centre  of  the  colony 
becomes  yellow. 

On  the  surface  of  gelatine  a  film 
develops,  which  in  a  few  days  turns 
yellow.  Old  cultures  have  a  pecu- 
liar smell. 

The  yellow  pigment  can  be  ex- 
tracted with  alcohol.  It  is  insoluble 
in  water,  and  decolorised  by  acids. 

They  were  obtained  from  human 
urine. 

Micrococcus  of  Forbes  (p. 
472). 

Micrococcus  plumpsus  (Brauti- 
gam). — Cocci  *8  /z  in  diam.,  in 
masses. 

Colonies  yellowish-white. 

Inoculated  in  the  depth  of  gela- 
tine long  delicate  acicular  processes 
shoot  out  from  the  needle  track 
and  on  the  free  surface. 

On  potato  the  growth  is  similar. 

They  were  isolated  from  water. 

Micrococcus  pneumoniae  crou- 
posae  (p.  236). 

Micrococcus  pyogenes  tenuis 
(Rosen  bach). — Cocci  irregular, 
somewhat  larger  than  staphylococci, 
and  with  much  less  tendency  to 
form  masses.  The  ends  colour 
deeply,  leaving  a  clear  space  in  the 
middle. 

Inoculated  in  the  depth  of  gela- 
tine a  slightly  opaque  growth  is 
formed. 

On  agar  a  thin  deposit  appears 
along  the  needle  track,  which  is 
almost  as  clear  as  glass. 

They  occur  in  the  pus  of  un- 
opened abscesses,  but  not  often,  as 
they  were  found  by  Rosenbach  only 
in  three  out  of  thirty-nine  cases. 


558 


DESCRIPTION   OF   SPECIES. 


Micrococcus  rpsettaceus  (Zim- 
mermann). — Cocci  '7  to  1  p.  in  diam., 
singly,  and  in  masses. 

Colonies  circular,  whitish,  or 
greyish-yellow. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  very  scanty  in 
the  track  of  the  needle,  but 
spreads  over  the  surface  as  a  grey 
rosette. 

On  agar  a  smooth  layer  with  den- 
tated  margin  is  formed. 

On  potato  the  growth  is  faintly 
yellowish. 

They  occur  in  water. 

Micrococcus  roseus  (Eisen- 
berg). — Cocci  forming  pink  colonies, 
and  a  rose-coloured  growth  on  the 
surface  of  nutrient  agar-agar. 

They  were  found  in  sputum  in  a 
case  of  influenza. 

Micrococcus  salivarius  septi- 
CUS  (Biondi). — Oval  cocci,  diplo- 
cocci,  in  chains  and  small  masses  ; 
capsulated. 

They  grow  best  on  acid  gelatine, 
or  in  an  atmosphere  of  carbonic 
acid. 

Colonies  are  small  and  circular, 
with  an  opalescent  centre  and  a 
transparent  margin,  with  sharply 
defined  outline.  In  the  interior  of 
the  colonies  there  is  an  appearance 
of  a  network. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  form  a  delicate  fila- 
ment and  white  dots  on  the  free 
surface. 

On  agar  the  cultures  should  be 
made  direct  from  the  blood.  The 
growth  appears  on  the  surface  and 
resembles  dewdrops. 

Broth  cultures  remain  clear. 

They  are  fatal  to  mice  in  twenty- 
four  to  seventy-two  hours,  and  to 
rabbits  in  fifteen  to  thirty  days, 
producing  septicaemia.  Attenuated 
cultures  are  said  to  give  immunity. 

They  were  found  in  the  saliva  of 
.healthy  and  diseased  persons. 

Micrococcus  stellatus  (Mas- 
chek). — Cocci  singly. 

Colonies  stellate. 

Inoculated  in  the  depth  of  gela- 
tine a  branching  growth  appears  in 
the  track  of  the  needle.  The  jelly 
becomes  brownish. 


On  potato  the  growth  is  brownish- 
yellow  and  shining. 

They  occur  in  water. 

Micrococcus  tetragenus 
(Gaffkey). — Cocci  about  1  /*  in 
diam.,  in  tetrads,  and  surrounded 
by  a  hyaline  capsule. 

Colonies  form  in  twenty-four  to 
forty-eight  hours  as  small  white 
dots,  which  are  finely  granular,  and 
have  a  vitreous  lustre  ;  when  they 
reach  the  surface  they  form  thick 
raised  masses. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  form  an  irregular 
white  growth,  especially  in  the 
upper  part  of  the  track  of  the 
needle. 

On  agar  the  colonies  occur  along 
the  needle  track,  and  are  white, 
round  and  circumscribed. 

On  potato  they  form  a  thick, 
slimy,  viscous  layer. 

White  mice  inoculated  with  a 
minute  quantity  of  a  pure-cultiva- 
tion die  in  from  two  to  ten  days, 
and  the  groups  of  the  characteristic 
tetrads  may  be  found  in  the  capil- 
laries throughout  the  body,  especi- 
ally in  the  spleen,  lung  and  kidney. 

Double  infection  can  be  produced 
by  inoculating  a  mouse  with  a 
pure  cultivation  of  Bacillus  an- 
thracis  two  or  three  days  after 
inoculation  with  Micrococcus  tetra- 
genus.  On  examination  after 
death,  the  capillaries  of  the  lungs, 
liver  and  kidney  are  filled  with 
both  anthrax  bacilli  and  masses  of 
tetrads  (Plate  V.,  Fig.  3). 

Micrococcus  tetragenus 
mobilis  ventriculi  (Mendoza).— 
Cocci  in  tetrads  ;  capsulated  ; 
motile. 

Colonies  circular,  whitish  and 
granular. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  on  the  free  surface 
only,  and  give  off  a  peculiar  odour. 

They  were  isolated  from  the 
stomach. 

Micrococcus  tetragenus  sub- 
flavus  (Von  Besser).— Cocci  singly, 
and  in  tetrads. 

They  do  not  grow  on  gelatine. 

Colonies  on  agar  are  brown  and 
irregular  in  contour. 


DESCRIPTION   OF   SPECIES. 


559 


On  the  surface  of  agar  the  cocci 
form  a  greyish-white  band,  which 
turns  brown  at  the  periphery,  and 
later  is  all  dark  or  orange-yellow. 

On  potato  the  growth  is  brown. 

They  occur  in  nasal  mucus. 

Micrococcus  tetragenus  ver- 
satilis  (Sternberg  and  Finlay). — 
Cocci  varying  in  size  from  '5  to  1*5 
yn,  in  tetrads  and  irregular  groups. 

Colonies  are  circular  and  lemon- 
yellow  in  colour. 

Inoculated  in  the  depth  of  gela- 
tine there  is  very  scanty  develop- 
ment along  the  line  of  puncture, 
and  the  gelatine  is  liquefied  in  the 
form  of  a  cup  near  the  surface. 
At  the  bottom  of  the  liquefied 
gelatine  a  viscid,  pale-yellow  mass 
accumulates. 

On  the  surface  of  agar  a  thick, 
viscid,  yellow  layer  is  formed  along 
the  line  of  inoculation,  which  gradu- 
ally extends  over  the  entire  sur- 
face. The  colour  varies  from  cream- 
yellow  to  lemon-yellow,  and  the 
surface  is  moist  and  shining. 

On  potato  there  is  a  similar 
growth. 

They  were  isolated  from  the 
skin  of  patients  suffering  from 
yellow  fever,  from  mosquitoes  after 
attacking  these  patients,  and  from 
the  air. 

Micrococcus  ureae  liquefaciens 
(Fliigge). — Cocci  spherical,  1*25  to 
2  fj,  in  diam.,  singly,  or  in  chains 
of  three  to  ten  elements,  or  in 
irregular  groups. 

Colonies  appear  in  two  days 
as  small  white  points.  They  have 
sharply  defined  edges  and  a  granular 
surface.  The  gelatine  gradually 
liquefies,  and  the  edges  of  the  colo- 
nies become  irregular. 

The  cocci  inoculated  in  the  depth 
of  gelatine  produce  a  continuous 
white  line  along  the  track  of 
the  needle.  Finally,  the  whole  of 
the  gelatine  liquefies,  and  appears 
as  a  whitish-turbid  fluid  with  a 
thick  whitish-yellow  deposit  at  the 
bottom. 

They  were  obtained  from  urine. 

Micro  co  ccusversicolor 
(Fliigge). — Small  cocci,  in  pairs 
and  in  masses. 


White  colonies  develop  in  twenty- 
four  hours  ;  after  two  days  they  are 
yellowish,  with  sharp  contours  of 
yellowish-green  colour,  and  finely 
granular.  The  superficial  colonies 
form  flat  deposits,  2'6  mm.  in  size, 
increasing  to  10  mm.  after  four  or 
five  days. 

On  the  surface  of  gelatine  the 
cocci  form  a  shining  layer  with  a 
greenish  or  bluish  shimmer  like 
mother-of-pearl . 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  composed  of 
spherical  yellowish  colonies,  and 
on  the  free  surface  they  form  an 
iridescent  film. 

They  occur  in  the  air. 

Micrococcus  violaceus  (Schro- 
ter). — Cocci  or  elliptical  cells,  de- 
scribed as  uniting  into  violet-blue 
gelatinous  spots,  which  again  unite 
to  form  larger  patches. 

The  colonies  on  gelatine  are  violet 
in  colour. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  scanty  in  the 
track  of  the  needle. 

On  the  surface  of  gelatine  they 
form  a  bluish-violet  layer,  and  the 
same  on  agar  and  potato. 

They  were  observed  on  boiled 
potatoes  exposed  to  the  air,  and  are 
also  found  in  water. 

Micrococcus  viticulosus  (Katz). 
— Oval  cocci  1  //  in  width,  and  1'2  /u, 
in  length,  in  masses,  but  without 
formation  of  much  gelatinous  ma- 
terial. 

The  superficial  colonies  are  quite 
different  in  appearance  from  the 
deep  colonies.  From  the  deep 
colonies  fine  hairlike  tendrils  are 
thrown  off  from  a  centre,  forming 
a  very  delicate  and  extensive  net- 
work. The  threads  are  found  to 
consist  of  zooglcea  masses,  irregular 
in  size,  arranged  like  strings  of 
beads.  The  colonies  which  are  ex- 
posed to  the  air  form  a  thin  layer 
of  muddy-white  gelatinous  sub- 
stance, which  rapidly  spreads,  some- 
times sending  out  hairlike  processes 
into  the  depth  of  the  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine a  delicate  feather-like  growth 
occurs  along  the  track  of  the  needle, 


560 


DESCRIPTION   OF   SPECIES. 


and  on  the  free  surface  they  pro- 
duce the  appearance  which  has  been 
described  in  colonies.  This  micro- 
organism is  exceedingly  rare.  It 
was  obtained  from  a  contaminated 
culture. 

Monas  Okenii. — Short  cylindrical 
cells,  5  n  wide,  8  to  15  p.  long,  with 
rounded  ends.  They  exhibit  lively 
movements,  each  end  being  provided 
with  a  flagellum  twice  as  long  as 
the  cell  itself.  They  have  pale-red 
cell-substance,  with  dark  grains. 

They  occur  in  stagnant  water. 

Monas  vinosa.—  Round  or  oval 
cells  of  about  2'5  p.  in  diam.,  often 
united  in  pairs.  Their  motion  is 
slow  and  tremulous,  and  the  cell- 
substance  is  pale-red  with  dark 
grains  interspersed.  Flagella  have 
not  been  observed. 

They  were  found  in  water  with 
decaying  vegetable  matter. 

Monas  Warmingii.— Cylindrical 
cells,  rounded  at  the  ends,  15  //,  long, 
5  to  8  /j.  broad.  They  are  possessed 
of  a  flagellum  at  each  end,  and 
exhibit  rapid,  irregular  movements. 
The  cell-substance  is  pale-red,  inter- 
spersed at  the  ends  with  dark-red 
grains. 

Myconostoc  gregarium  (Cohn). 
— The  threads  are  very  thin,  colour- 
less, unarticulated,  but  fall  apart 
into  short  cylindrical  links  when 
dried. 

They  form  gelatinous  masses, 
10  to  17  n  in  diam.,  singly  or 
heaped  into  slimy  drops  011  water 
in  which  algae  are  decomposing. 

Nitromonas  of  Winogradsky.— 
Very  short  rods,  -9  to  1  p,  in  width, 
I'l  to  T8  p.  in  length.  Singly,  in 
masses,  and  in  very  short  chains. 

They  can  be  cultivated  in  silica- 


'hey  are  active  agents  of  nitrifi- 
cation. 

They  were  obtained  from  the  soil. 

Pediococcus  acidi  lactici 
(Lindner). — Cocci  '6  to  1  ^  in  diam., 
singly,  in  pairs,  and  tetrads. 

Colonies  colourless. 

On  the  surface  of  agar  the  cocci 
form  a  colourless  layer. 

On  potato  the  growth  is  almost 
invisible. 


The  cocci  produce  lactic  acid  in 
solutions  containing  sugar. 

They  occur  in  hay  infusion  and 
malt. 

Pediococcus  cerevisiae  (Balcke). 
— Cocci  singly,  in  pairs,  and  tetrads. 

Colonies  at  first  colourless,  later 
yellowish-brown. 

Inoculated  in  the  depth  of  gela- 
tine a  greyish-white  filament 
occurs  in  the  track  of  the  needle, 
and  a  white  layer  on  the  free 
surface. 

On  agar  the  growth  is  transparent 
and  iridescent,  and  on  potato  almost 
invisible. 

They  were  isolated  from  the  air 
of  a  brewery. 

Pneuinobacillus  liquefaciens 
bovis  (p.  242). 

Proteus  capsulatus  septicus 
(Banti). — Rods  isolated  from  a  case 
of  septica3mia,  and  identical  with 
Proteus  hominis  capsulatus. 

Proteus  hominis  capsulatus 
(p.  224). 

Proteus  in  gangrene  of  the 
lung  (Babes). — Rods  -8  to  1-5  /* 
thick,  irregular  in  form,  and  fila- 
ments with  irregular  enlargements. 

Colonies  whitish  and  transparent, 
with  ramifications  extending  over 
the  gelatine. 

In  the  depth  of  gelatine  a  growth 
occurs  along  the  track  of  the 
needle,  and  a  ramifying  growth  on 
the  free  surface. 

On  agar  the  growth  is  slightly 
yellowish. 

On  potato  the  growth  is  brownish. 

They  are  extremely  pathogenic 
in  mice  and  guinea-pigs. 

They  were  isolated  from  a  case 
of  gangrene  of  the  lung. 

Proteus  microsepticus  (Kar- 
linski). — Cocci,  rods  and  filaments 
in  morphology,  and  cultures  re- 
sembling Proteus  vulgaris. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the 
track  of  the  needle,  forming  a 
funnel  with  cloudy  contents,  and 
in  a  few  days  the  whole,  of  the 
gelatine  is  liquid. 

They  produce  a  general  infection 
in  mice,  and  death  in  twenty-four 
hours,  and  occasionally  death  in 


DESCRIFHON   OF   SPECIES. 


561 


rabbits,   and  local   suppuration  in 
guinea-pigs  and  white  rats. 

They  were  isolated  from  pus  in 
a  fatal  case  of  puerperal  pyaemia. 


FIG.  221. — PROTEUS  MIRABILIS  :  SWARMING 
ISLANDS  ON  THE  SURFACE  OF  GELATINE,  x  285 
(HAUSER). 


Proteus  mirabilis.— Cocci  -4  /* 
to  '9  /LI.  They  occur  singly  and  in 
zoogloea,  and  sometimes  in  tetrads, 
pairs,  chains,  or  as  short  rods  in 
twos  resembling  Bacterium  termo — 


concentric   circles,   which    in   time 
liquefies     the    medium.        Similar 
movements  are  observed  in  capsule- 
cultivations  as  in  Proteus  vulgaris. 
They  were  isolated  from 
putrid  meat  infusion. 

Proteus  septic  us 
(Babes).— Kods  -4  /*  in 
width,  and  filamentous 
forms. 

Colonies  rapidly  liquefy 
^^s**    the  gelatine. 

Inoculated  in  the  depth 
/          of  gelatine  the  bacilli  form 
/  a   turbid   funnel,  or  com- 

pletely liquefy  the  gelatine 
in  twenty-four  hours. 

On  agar  the  growth  is 
reticulated. 

On  potato  brownish - 
white. 

Cultures  have  an  un- 
pleasant odour. 

They  are  pathogenic  in 
mice. 

They  were  isolated  from 
the  organs  in  a  case  of 
human  septicaemia. 

Proteus  sulfureus  (Lin- 
denborn). — Rods  -8  /*  in 
width,  varying  in  length, 
and  long  filaments. 

They  correspend  in  mor- 
phology and  cultures  with 
Proteus  vulgaris. 

They  produce   sulphuretted  hy- 
drogen in  cultures. 

They  were  isolated  from  water. 
Proteus    vulgaris    (Hauser).— 


*%•» 


£  -i-^^sf*        *     »  x    *<r       ><7^ 

%  ""^     Si  /J/      ^« ^o/t  \' 

£>.*flPvT.^ 


FIG.  222.— PROTEUS  MIRABILIS.:  INVOLUTION  FORMS,   x  524  (HAUSER). 


in  fact,  in  all  conceivable  transition 
forms. 

Cultivated   on  nutrient  gelatine 
they  form  a  thick  whitish  layer  in 


Rods  varying  in  size  ;  some  mea- 
sure 4  /n  in  length,  and  are  almost 
as  broad  as  long,  and  others  vary 
from  -94  to  1'25  p,  long  and  '42  to 

36 


562 


DESCRIPTION   OF  SPECIES. 


•63    fj.    wide.      They    are  actively 
motile. 

Cultivated  on  nutrient  gelatine 
they  convert  it  into  a  turbid,  grey- 
ish-white liquid.  If  cultivated  in 
a  capsule  containing  5  per  cent,  of 
nutrient  gelatine,  a  few  hours  after 
inoculation  the  most  characteristic 
movements  of  the  individual  bacilli 


FIG.   223. — PROTEUS    VULGARIS,     FROM 
SURFACE  OF  NUTRIENT  GELATINE,    x 
(HAUSER). 

are  observed  on  the  surface  of  the 
nutrient  gelatine,  although  at  this 
early  stage  no  superficial  liquefac- 
tion can  be  detected.  Probably  the 
movements  depend  upon  the  exist- 
ence of  a  thin  liquid  layer,  as  they 
are  not  observed  if  the  nutrient 
medium  contains  10  per  cent,  of 
gelatine. 

They  were  isolated  from  putrid 
meat  infusion. 

Proteus  Zenkeri.— Cocci  -4  /*,  in 
twos  like  Bacterium  termo,  and 
short  rods  1  -65  /*  long. 

Cultivated  on  nutrient  gelatine 
no  liquefaction  results,  but  a  thick 
whitish-grey  layer  is  formed.  The 


bacilli  are  motile,  and  the  same 
phenomena  are  observed  on  the 
solid  medium  as  in  Proteus  vulgaris. 
In  cover-glass  impressions  most 
varied  groupings  of  the  bacilli  are 
seen,  and  also  developmental  and 
involution-forms. 

They  were  isolated  from  putrid 
meat  infusion. 

Pseudo-diphtheritic  bacil- 
lus (p.  335). 

Pseudo-diplococcus  pneu- 
nioniae  (Bonome).— Oval  cocci 
in  pairs  and  short  chains  ;  cap- 
sulated. 

Inoculated  in  the  depth  of 
gelatine  small  colonies  develop 
in  the  track  of  the  needle  in 
twenty-four  hours. 

On  agar  there  is  a  scanty, 
moist  growth. 

On  potato  an  almost  in- 
visible film. 

In  broth  the  cocci  grow 
rapidly,  and  the  cultures  give 
off  a  peculiar  odour. 

They  produce  septicaemia  in 
mice,  guinea-pigs  and  rabbits. 
This       micro-organism       is 
probably     a    variety    of     the 
pneumococcus. 

They  were  isolated  from  a 
fatal  case  of  cerebro-spinal 
meningitis. 

Rhabdomonas       rosea.  — 
Spindle-form  rods,  3'8  to  5  p 
THE    broad,  20  to  30  ^  long.     They 
3    exhibit  slow,  trembling  move- 
ments, having  at  each  end  of 
the    cell   a    flagellum.       The 
cell-substance   is  very    pale,    with 
dark  grains  interspersed. 

They  occur  in  brackish  water. 
S  a  r  c  i  n  a   alba. — Small  cocci. 
They  form  small  white  colonies  on 
nutrient  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  slightly  along  the 
needle  track,  but  are  heaped  up  on 
the  surface  without  liquefying  the 
gelatine. 

They  are  present  in  the  air. 
S  a  r  c  i  n  a    aurantiaca.— Cocci 
singly,  in  pairs,  in  tetrads,  and  in 
packets. 

Colonies  orange-yellow. 
Inoculated     in     the     depth  i  of 


DESCRIPTION    OF   SPECIES. 


563 


gelatine  they  slowly  liquefy  it  along 
the  whole  needle  track,  and  form 
on  the  surface  an  orange-yellow 
growth.  On  potatoes  they  slowly 
develop  the  same  pigment. 

Sarcina  Candida  (Reinke).— 
Cocci  1-5  to  1-7  /x  in  diam.,  singly, 
in  pairs,  and  in  tetrads. 

Colonies  are  circular  and  shining, 
white,  and  later  yellowish. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  quickly  takes  place 
along  the  track  of  the  needle. 

On  the  surface  of  agar  a  white, 
moist  layer  develops. 

They  were  found  in  the  air  of 
breweries. 

Sarcina  flava(De  Bary).— Small 
cocci  in  packets. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  liquefaction. 

On  agar  they  form  a  yellow 
layer. 

On  potato  the  growth  is  limited 
and  yellow. 

They  were  isolated  from  beer. 

Sarcina  hyalina  (Kiitzing). — 
Cocci  round,  2'5  ^  in  diam.,  almost 
colourless.  United  in  families  of 
4  to  24  cells,  reaching  15  /u  in  diam. 

They  occur  in  marshes. 

Sarcina  intestinalis  (Zopf).— 
Cocci  in  groups  of  four  or  eight. 
Yery  regular  in  form  ;  never  in 
the  large  packets  which  occur  in 
Sarcina  ventriculi. 

They  are  found  in  the  intestinal 
canal,  especially  the  caecum,  of 
poultry,  particularly  fowls  and 
turkeys. 

Sarcina  litoralis  (Oersted).— 
Cocci  1-2  to  2  fj.  in  diam.,  bound 
together  in  4  to  8  families,  which, 
in  their  turn,  may  unite  and  in- 
clude as  many  as  64  tetrads. 
Plasma  colourless  ;  in  each  cell 
1  to  4  sulphur  granules. 

They  were  found  in  sea  water 
containing  putrefying  matter. 

Sarcina  lutea  (Schroter). — 
Cocci  singly,  in  pairs,  tetrads  and 
packets.  A  single  individual  in  a 
tetrad  may  be  divided  into  two,  or 
into  four,  so  that  a  tetrad  within  a 
tetrad  results. 

Colonies  are  round,  slightly 
granular  in  appearance,  and  yellow. 


Inoculated  in  the  depth  of 
gelatine  they  grow  rapidly  ;  the 
gelatine  becomes  liquefied,  and  the 
yellow  growth  sinks  to  the  bottom 
of  the  tube. 


as* 


FIG.  224.— SARCINA  x  600  (FLUGGE. 


Cultivated  in  agar  they  form  a 
colourless  growth  along  the  track 
of  the  needle,  and  a  bright  canary- 
yellow  layer  upon  the  surface. 

On  potato  they  form  a  yellow 
layer. 

They  are  present  in  air. 

Sarcina  mobilis  (Maurea).— 
Cocci  1-5  p.  in  diam.,  in  pairs,  and 
in  tetrads.  They  are  motile. 

Colonies,  at  first  white,  become 
brick-red. 

Inoculated  in  the  depth  of  gela- 
tine, there  is,  after  several  days,  a 
slight  growth  along  the  track  of 
the  needle,  and  a  patch  of  growth 
on  the  free  surface  which  gradually 
turns  red.  In  about  two  weeks 
liquefaction  produces  a  funnel- 
shaped  appearance  ;  later  the  lique- 
faction extends  to  the  sides  of  the 
test-tube. 

In  broth  turbidity  is  produced, 
and  a  yellowish-red  deposit. 

On  agar  the  growth,  at  first  white, 
changes  to  a  brick-red  colour. 

There  is  no  growth  on  potato, 
and  milk  is  not  coagulated. 

They  were  isolated  from  ascitic 
fluid. 

Sarcina  pulmonum  (Hauser).— 
Cocci  from  1  to  1*5  ft  in  diam.,  in 
tetrads  and  packets. 

Colonies  white  and  small.  They 
are  coarsely  granular. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  is  scanty  in  the 
track  of  the  needle,  but  on  the  free 
surface  there  is  a  circular,  well- 
defined,  translucent  patch,  which 


564 


DESCRIPTION   OF   SPECIES. 


later  becomes  greyish -brown,  shin- 
ing, wrinkled  and  irregular. 

On  potato  the  growth  is  very 
slight  and  limited. 

They  cause  ammoniacal  decom- 
position of  urine. 

They  were  isolated  from  phthisi- 
cal sputum. 

Sarcina  Reitenbachii  (Caspary). 
— Cocci  about  1'5  to  2 '5  \i  in  diam., 
at  the  time  of  division  lengthened 
to  4  \i.  Mostly  united  together 
from  4  to  8  in  number  ;  occasion- 
ally 16  or  more.  Colourless  cell- 
wall,  lined  with  rose -red  layer  of 
plasma. 

They  were  found  on  rotting 
water-plants. 

Sarcina  rosea  (Schroter). — 
Large  cocci,  in  packets. 

Inoculated  in  the  depth  of  gela- 
tine, liquefaction  quickly  takes 
place,  and  cultures  after  a  time 
have  a  reddish  colour. 

On  agar  the  growth  is  slow  and 
limited. 

On  potato  the  growth  is  abundant 
and  of  a  bright-red  colour. 

In  broth  they  produce  turbidity, 
and  a  red  deposit. 

They  occur  in  the  air. 

Sarcina  urinse  (Welcker).— Very 
small  cocci,  1'2  /z  in  diam.,  united 
in  families  of  8  to  64.  They  were 
observed  in  urine. 

Sarcina  yentriculi  (Goodsir).— 
Cocci  reaching  4  /*  in  diam.,  united 
in  groups  of  four,  or  multiples  of 
four,  producing  cubes  or  packets 
with  rounded-off  corners.  Contents 
of  the  cells  are  greenish  or  yellow- 
ish-red. 

Colonies  are  round  and  yellowish. 

A  yellowish  growth  forms  on  the 
surface  of  oblique  gelatine  without 
liquefaction. 

On  potato  they  form  a  yellow 
growth,  and  on  serum  also. 

They  grow  well  in  hay-infusion, 
forming  brownish  scales  and  a  simi- 
larly-coloured deposit. 

They  occur  in  the  stomach  of  man 
and  animals  in  health  and  disease, 
and  were  first  detected  in  vomit. 

Sphaerotilus  natans.— Cells  4  to 
9  /z  long,  and  3  /x  thick,  united  in 
a  gelatinous  sheath  to  form  threads. 


The  cells  comprise  rods  and  cocci- 
forms  ;  the  cocci  are  set  free,  and 
develop  into  rods,  which  again  form 
threads.  In  the  last  a  false  branch- 
ing has  been  observed.  The  plasma 
of  the  cells  break  up  into  minute, 
strongly-refractive  portions,  which 
develop  into  round  spores,  at  first 
of  a  red  and  afterwards  a  brown 
colour. 

They  occur  in  stagnant  and  flow- 
ing water  contaminated  with  or- 
ganic matter,  and  form  floating 
flakes  of  a  white,  yellow,  rust-red, 
or  yellow-brown  colour. 

Spirillum  amyliferum  (Van 
Tieghem). — Filaments  6ju  in  length 
and  1'4  to  1'5  /*  in  width  ;  with 
from  2  to  4  screw  curves. 

They  act  as  a  strong  ferment  in 
the  absence  of  air. 
They  occur  in  water. 
Spirillum  anserum  (Sakharoff). 
— Spirilla    resembling    the     Spiro- 
chae ta    0 bermeieri.     Extremely 
motile. 

They  have  not  been  cultivated 
artificially. 

Blood  from  diseased  geese,  con- 
taining the  spirilla,  produces  the 
disease  when  inoculated  in  healthy 
birds.  The  geese  suffer  from 
diarrhoea,  and  die  in  about  a  week. 
They  were  found  in  the  blood  of 
geese  suffering  from  an  epidemic 
form  of  septicaBmia  prevailing  at 
some  of  the  stations  on  the  Trans- 
caucasian  Railway. 

Spirillum  attenuatum  (Warm- 
ing).— Threads  much  attenuated  at 
the  ends,  which  consist  usually  of 
three  spirals.  The  middle  spiral  is 
about  11  p,  high  and  6  p.  in  diam., 
and  the  end  ones  10  ft  high  and  2  p. 
in  diam. 

They  are  found  in  brackish  water. 
Spirillum  aureum  (Weibel). — 
Curved  rods  with  blunt  ends,  spi- 
rilla and  spirilliform  filaments,  and 
involution  forms. 

Colonies  are  circular  and  golden- 
yellow. 

Inoculated  in  the  depth  of  gela- 
tine, a  finely  granular  growth  forms 
in  the  track  of  the  needle,  and  a 
yellow-ochre  prominent  mass  on 
the  free  surface. 


DESCRIFHON   OF   SPECIES. 


565 


On  agar  a  greyish  growth  ex- 
tends over  the  surface,  and  later 
prominent  yellowish  heaps  make 
their  appearance. 

On  potato  there  is  an  abundant 
growth  of  a  golden-yellow  colour. 

They  were  found  in  sewage 
mud. 

Spirillum  cholerae  Asiaticse 
(p.  301). 

Spirillum  choleroides  (Buj  wid). 
— Curved  rods  very  similar  mor- 
phologically and  in  cultures  to 
Koch's  comma-bacilli. 

They  were  isolated  from  river 
water. 

Spirillum  choleroides  (Orlow- 
sld). — Curved  rods  very  similar  to 
Koch's  comma-bacilli. 

They  were  found  in  well  water. 

Spirillum  concentricum 
(Kitasato). — Short  spirilla,  and 
spirilliform  filaments. 

Colonies  are  circular,  and  com- 
posed of  concentric  rings  alternately 
opaque  and  transparent. 

Inoculated  in  the  depth  of  gela- 
tine there  is  only  a  little  growth 
in  the  track  of  the  needle,  and  a 
cloudy  growth  on  the  surface 
extending  into  the  jelly. 

On  agar  the  growth  is  extremely 
adherent. 

In  broth  they  produce  turbidity, 
which  disappears  after  a  time  ;  and 
there  is  a  slimy  deposit  at  the 
bottom  of  the  tube. 

They  were  found  in  putrefying 
blood. 

Spirillum  dentium  (Miller).— 
Spirals  10  to  20  p,  in  length,  pointed 
at  the  ends. 

They  have  not  been  cultivated. 

They  occur  in  the  deposit  on  the 
teeth,  and  in  company  with  Lepto- 
thrix  buccalis  in  carious  teeth. 

Spirillum  flavescens  (Weibel). 
— Commas  thicker  than  those  found 
in  Asiatic  cholera,  spirilla,  and 
spirilliform  filaments. 

Colonies  yellowish. 

Inoculated  in  the  depth  of  gela- 
tine a  finely  granular  filament 
develops  in  the  track  of  the  needle, 
and  on  the  free  surface  a  pale- 
yellow  patch. 

On    agar    the    growth,    at    first 


greyish- white,  becomes  yellow,  and 
forms  a  thick  layer. 

On  potato  the  growth  is  abun- 
dant, and  similar  in  colour. 

They  were  found  in  sewage 
mud. 

Spirillum  flavum  (Weibel).— 
Spirilla  morphologically  identical 
with  spirillum  aureum. 

Colonies  on  gelatine  are  pale- 
yellow,  and  later  the  colour  is  more 
intense. 

On  agar  and  potato  they  form  a 
layer  the  colour  of  yellow-ochre. 

They  were  isolated  from  sewage 
mud. 

Spirillum  leucomelaneum 
(Koch). — Dark  and  glass-like  spaces 
alternate  in  the  spirillum,  resulting 
from  a  regular  arrangement  of  the 
dark  granular  contents.  A  rare 
form  observed  in  water  covering 
rotting  algaB. 

Spirillum  linguae  (Weibel). — 
Curved  rods,  spirilla,  and  spirilli- 
form filaments,  and  involution 
forms. 

Colonies  are  composed  of  inter- 
lacing filaments,  and  offshoots 
extend  into  the  surrounding  gela- 
tine. 

Inoculated  in  the  depth  of  gela- 
tine a  delicate  growth  occurs  in  the 
needle  track. 

On  agar  the  growth  is  whitish 
and  granular. 

In  broth  a  cloudiness  is  produced, 
as  well  as  a  flocculent  deposit. 

They  are  especially  distinguished 
from  other  spirilla  described  by 
Weibel  by  their  staining  by  Gram's 
method. 

They  were  isolated  from  the 
tongue. 

Spirillum  marinum  (Russell). 
— Curved  rods,  and  spiral  filaments. 

Colonies  circular,  granular  and 
striated  ;  later,  flocculent  masses 
float  in  liquefied  areas. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the 
track  of  the  needle,  and  a  mem- 
brane forms  on  the  surface  of  the 
cloudy  liquid. 

On  agar  the  growth  is  yellowish 
and  abundant. 

On   potato   a  thick,   waxy  mass 


366 


DESCRIPTION   OF   SPECIES. 


develops,  and  extends  over  the 
surface. 

Broth  made  with  sea- water  be- 
comes rapidly  turbid. 

They  were  obtained  from  sea- 
water. 

Spirillum  Metchnikovi  (p.  373). 

Spirillum  nasale  (Weibel).— 
Large  curved  rods  and  spirilliform 
filaments.  Non-motile. 

Colonies  circular,  finely  granular, 
and  brownish-yellow. 

Inoculated  in  the  depth  of  gela- 
tine a  delicate  growth  develops  in 
the  track  of  the  needle. 

On  the  surface  of  agar  they  form 
a  whitish  slimy  film. 

They  occur  in  nasal  mucus. 

Spirillum  Obermeieri  (p.  258). 

Spirillum  of  Finkler  and 
Prior  (p.  258). 

Spirillum  of  Gimther  (Vibrio 
aqudtilis) . — Curved  rods,  very  simi- 
lar to  Koch's  comma-bacilli,  but 
there  is  no  growth  on  potato. 

They  occur  in  water. 

Spirillum  of  Miller.— Curved 
rods,  singly,  in  pairs,  and  spiral 
filaments. 

They  liquefy  gelatine. 

They  were  isolated  from  carious 
teeth. 

Spirillum  of  Neisser  (Vibrio 
berolinensis). — Similar  to  Koch's 
comma-bacillus,  but  smaller. 

Colonies  colourless,  granular  and 
transparent,  liquefying  the  gelatine 
much  more  slowly  than  Koch's 
comma-bacillus. 

Inoculated  in  the  depth  of  gela- 
tine they  produce  a  growth  similar 
to  that  of  Koch's  comma-bacilli, 
but  much  more  slowly  in  milk. 

In  broth  they  grow  abundantly. 

They  were  found  in  water. 

Spirillum  of  Renon.— Curved 
rods  longer  and  broader  than  the 
comma-bacilli  of  Koch. 

Colonies  yellowish,  with  dark 
nucleus. 

In  gelatine  the  cultures  resemble 
those  of  Koch's  comma-bacillus. 

On  agar  the  growth  is  white  and 
abundant. 

They  cause  turbidity  in  broth. 

They  were  isolated  from  impure 
water  from  a  well. 


Spirillum  of  Smith  (vide 
Spirillum  suis). 

Spirillum  of  Weib-el.— Curved 
rods  resembling  Koch's  comma- 
bacilli,  morphologically,  and  in 
cultures  on  jelly.  The  gelatine  is 
more  quickly  liquefied. 

There  is  no  growth  on  potato. 

They  occur  in  well  water. 

Spirillum  plicatile  (Ehrenberg). 
— Thin  threads  2'25  fj,  in  breadth, 
110  to  125  /u,  long,  occurring  also  in 
spirulinar  forms.  The  threads  have 
primary  and  secondary  windings  ; 
the  former  are  in  each  example  of 
equal  size,  but  the  latter  are  often 


FIG  225.—  SPIKOCH.ETA  PLICATILE. 

irregular.  Their  ends  are  cut  off 
bluntly,  and  they  exhibit  rapid 
movement. 

They  occur  abundantly  in  marsh- 
water  in  summer,  and  can  be  ob- 
tained by  allowing  algae  to  decom- 
pose in  water.  On  cultivation  the 
threads  break  up  into  long  rods, 
short  rods,  and,  finally,  cocci.  This 
change  is  rendered  visible  by  making 
cover-glass  preparations,  and  stain- 
ing with  aniline  dyes. 

Spirillum  rosaceum  (Klein). — 
Resembles  Spirillum  undula,  but  is 
reddish  in  colour  ;  the  colouring- 
matter  is  insoluble  in  water,  alcohol 
or  chloroform. 

Spirillum  Ros  enbergii.— 
Threads  with  1  to  1£  windings,  4 


DESCRIPTION   OF   SPECIES. 


567 


to  12  p  long,  1-5  to  2-6  p  thick. 
They  are  colourless,  but  the  con- 
tents include  strongly  refractive 
sulphur  granules.  Also  spirals  6  to 
7'5  \i  in  height,  which  are  actively 
motile. 

They  were  found  in  brackish 
water. 

Spirillum  rubrum  (Esmarck). 
— Curved  rods,  spirilla  and  spiro- 
chetae.  They  are  actively  motile. 

The  growth  on  artificial  media  is 
extremely  slow. 

Inoculated  in  the  depth  of  gela- 
tine they  grow  along  the  track  of 
the  needle,  forming  a  filament  of  a 
wine-red  colour,  without  causing 
liquefaction  ;  and  on  the  free  sur- 
face the  growth  is  colourless. 

In  broth  long  spirillar  threads 
are  formed. 

They  were  isolated  from  the 
putrid  tissues  of  a  mouse. 

Spirillum  rufum  (Perty).— 
Filaments  from  8  to  16  /z  in  length, 
with  1^  to  4  screw  curves  ;  non- 
segmented  ;  chiefly  motile  ;  tinged 
with  red. 

They  form  rose  or  dark  red  spots 
on  the  sides  of  wells. 

Spirillum  sanguineum  (Cohn). 
— Threads  3  \L  and  more  in  thick- 
ness, with  2  to  2£  spirals,  each  9  to 
12  /u.  high.  The  ends  are  provided 
with  flagella.  Their  colour  is  due 
to  the  presence  of  reddish  granules 
contained  in  the  cells. 

They  were  observed  in  brackish 
water  containing  putrefying  sub- 
stances. (Vide  Beggiatoa  roseo- 
persicina. ) 

Spirillum  saprophiles. — (I.) 
Curved  rods  with  pointed  ends,  -6  /L* 
in  width,  3  \JL  in  length  ;  spirilla, 
spirilliform  filaments,  and  involu- 
tion forms. 

Colonies  yellowish  or  greenish- 
yellow. 

Inoculated  in  the  depth  of  gela- 
tine a  white  growth  forms  in  the 
track  of  the  needle,  later  becoming 
yellowish  ;  and  on  the  free  surface 
there  is  a  white  growth,  and 
beyond  this  a  transparent  film 
spreads  over  the  jelly. 

On  agar  the  growthi  s  creamy, 
and  the  jelly  clouded  beneath  it. 


On  potato  the  growth  is  slimy 
and  yellowish  or  dark-brown  in 
colour. 

They  were  obtained  from  sewage 


FIG.  226.— COMMA-BACILLI  IN   WATER 

CONTAMINATED   WITH  SEWAGE. 


mud  and  decomposing  hay  infu- 
sion. 

(II.)  Curved  rods  about  2  p  in 
length,  with  blunt  ends  and  in  pairs. 
Extremely  motile. 

Colonies  circular  and  yellowish- 
brown. 

Inoculated  in  the  depth  of  gela- 
tine a  white  growth  develops  in 
the  track  of  the  needle,  and  later 
becomes  yellowish-red  ;  on  the  free 
surface  a  white  patch  forms,  sur- 
rounded by  a  transparent  film. 

In  the  depth  of  agar  there  is  no 
growth  in  the  track  of  the  needle, 
but  a  yellowish-white  patch  on 
the  free  surface  adherent  to  the 

jelly. 

On  potato  the  growth  is  also 
adherent,  and  in  appearance  shining 
and  brownish-green. 

They  were  isolated  from  decom- 
posing hay  infusion. 

(III.)  Curved  rods,  spirilla,  and 
spirilliform  filaments,  and  involu- 
tion forms. 

Colonies  are  circular,  granular, 
and  with  irregular  margin  ;  yellow 
in  the  centre,  and  white  at  the 
periphery. 

Inoculated  in  the  depth  of  gela- 
tine a  white  growth  develops  in 
the  track  of  the  needle,  and  on  the 
surface,  without  producing  lique- 
faction. 

On  the  surface  of  agar  the  growth 
is  white. 

On  potato  the  growth  is  distinctly 
brown. 


568 


DESCRIPTION   OF   SPECIES. 


They  were  isolated  from  sewage 
mud. 

Spirillum  serpens  (Miiller).— 
Long  spirilliform  filaments  ;  often 
collected  in  masses. 

They  were  observed  in  vegetable 
infusions  and  stagnant  water. 

Spirillum  sputigenum  (Lewis). 
— Curved  rods,  very  similar  to  the 
comma-bacilli  of  Koch  ;  but  many 
observers  having  failed  in  repeated 


,>\vvVv» 

<&?.<&• 


FIG.  227. — COMMA-BACILLI     OP     THE 
MOUTH,    x  700  (VAN  ERMENGEM). 

attempts  to  cultivate  them,  main- 
tain that  they  are  biologically  dis- 
tinct from  those  associated  with 
Asiatic  cholera.  Klein  asserts  that 
they  can  be  cultivated  in  an  acid 
gelatine,  and  that  they  are  iden- 
tical with  Koch's  comma-bacilli  in 
their  mode  of  growth.  They 
occur  with  other  bacteria  in  saliva 
and  in  scrapings  from  carious 
teeth. 

Spirillum  suis  (Smith).— Com- 
mas and  spirilla. 

Colonies  in  gelatine  are  circular, 
granular  and  brownish,  and  later 
appear  to  be  composed  of  concen- 
tric rings.  The  gelatine  is  not 
liquefied. 

Broth  with  1  per  cent,  of  peptone 
becomes  in  a  few  days  clouded. 

On  potato  they  develop  a  thin 
yellowish  layer. 

The  commas  are  said  to  be  slightly 
larger  than  those  obtained  from 
Asiatic  cholera,  and  are  not 
pathogenic. 

They  were  obtained  from  the 
large  intestine  in  swine. 

Spirillum  tenue.— Very  thin 
threads,  with  at  least  1J,  usually 
2  to  5  spirals.  Height  of  a  single 
screw  is  2  to  3  /u,  and  the  length  of 
spiral  therefore  4  to  15  p..  They 
are  very  swiftly  motile. 


They  often  occur  in  dense  felted 
swarms  in  vegetable  infusions. 

Spirillum  tyrogenum  (Deneke). 
— Curved  rods,  slightly  smaller 
than  Koch's  comma-bacilli,  with 
a  great  tendency  to  form  long 
spirillar  threads  (Fig.  228). 


FIG.  228.  —  DENEKE'S    COMMA-BACILLI 
FROM  CHEESE,  x  700  '.F 


Colonies  on  plate-cultivations 
are  sharply  defined  and  of  a 
greenish-brown  colour.  After  a 
time  they  liquefy  the  gelatine,  but 
the  liquefaction  is  much  more 
marked  than  in  colonies  of  Koch's 
commas  of  the  same  age,  though 
not  so  rapid  as  in  the  case  of  the 
commas  of  cholera  nostras. 

Inoculated  in  the  depth  of  nu- 
trient gelatine  a  turbid  liquefaction 
occurs  along  the  needle  track,  and 
on  the  surface  of  nutrient  agar-agar 
a  yellowish-white  layer  develops. 

Inoculation  of  potatoes  gives  no 
result. 

Administration  of  the  bacilli  by 
the  mouth,  in  the  manner  employed 
for  testing  the  pathogenic  effect  of 
Koch's  bacilli,  produced  a  fatal 
result  in  a  few  cases  ;  on  the  other 
hand,  injection  into  the  duodenum 
failed  entirely.  The  pathogenic 
properties  may  be  therefore  con- 
sidered as  not  yet  established. 

They  were  isolated  from  old 
cheese. 

Spirillum  undula.—  Threads 
1-1  to  1-4  fi  thick,  9  to  12  ^  long  ; 
spirals  4'5  fi  high  ;  each  thread  has 
1|  to  3  spirals.  They  are  actively 
motile,  and  possess  a  flagellum  at 
each  end. 

They  occur  in  various  infusions. 

Spirillum  vulutans(Ehrenberg). 
—Threads  1-5  to  2  p,  thick,  25  to 
30  p  long,  tapering  towards  their 


DESCRIPTION   OF   SPECIES. 


569 


extremities,  which  are  rounded  off. 
They  possess  dark  granular  con- 
tents. Each  thread  has  2£  to  3£ 
windings  or  spirals,  whose  height  is 
1»  to  13  p.  They  have  a  flagellum 
at  each  end,  and  are  sometimes 
motile,  sometimes  not. 

They  are  found  in  the  water  of 
marshes  and  in  various  infusions. 

Spirompnas  Cohnii.— Colourless 
cells,  consisting  of  1^  spirals,  with 
both  ends  acutely  pointed  and  pro- 
vided with  a  flagellum.  Breadth 
of  the  cells,  1-2  to  4  /u. 

They  occur  in  water  containing 
decomposing  matter. 

Spiromonas  volubilis  (Perty). 
— Colourless,  transparent  cells,  15 
to  18  p  long.  Rapidly  motile,  and 
revolving  round  a  longitudinal 
axis. 

They  occur  in  marsh-water  and 
putrefying  infusions. 

Staphylococcus  pyogenes  albus 
(p.  178). 

Staphylococcus  pyogenes 
aureus  (p.  176). 

Staphylococcus  pyogenes  cit- 
reus  (p.  178). 

Staphylococcus  pyosepticus 
(Heucourt  and  Richet). — Cocci 
identical  with  Staphylococcus  pyo- 
genes aureus. 

Subcutaneous  injection  causes  in 
rabbits  intense  oedema,  and  death 
in  twenty-four  hours. 

They  were  isolated  from  pus 
from  an  abscess  in  a  dog. 

Staphylococcus  salivarius  pyo- 
genes (Biondi). — Cocci  '3  to  -5  /* 
in  diam.,  singly  and  in  masses. 

Colonies  white  and  opalescent, 
producing  liquefaction. 

Inoculated  in  the  depth  of  gela- 
tine the  growth  appears  in  the 
track  of  the  needle,  and  is  followed 
by  liquefaction. 

On  agar  the  growth  is  orange- 
yellow. 

The  cocci  produce  local  suppura- 
tion when  inoculated  in  animals. 

They  were  isolated  from  an 
abscess  in  a  guinea-pig  following 
subcutaneous  injection  of  saliva. 

This  coccus  is  probably  identical 
with  Staphylococcus  pyogenes 
aureus. 


Staphylococcus  viridis  flaves- 
cens  (Guttmann). — Cocci  singly,  in 
pairs  and  masses  ;  morphologically 
agreeing  with  Staphylococcus  pyo- 
genes aureus. 

Colonies  are  greenish-yellow. 

Inoculated  in  the  depth  of  gela- 
tine a  filament  forms  composed  of 
greyish  colonies. 

On  agar  the  growth  is  greenish- 
yellow. 

They  grow  well  on  potato. 

They  were  isolated  from  the 
vesicles  of  chicken-pox. 

Streptococcus  acidi  lactici 
(Grotenfeld). — Oval  cocci  -5  to  1  /n 
long,  '3  to  '6  //,  in  width,  and  long 
chains.  They  are  partially  anaerobic. 

Colonies  are  circular  and  white. 

Inoculated  in  the  depth  of  gela- 
tine a  growth  occurs  only  in  the 
track  of  the  needle. 

Milk  is  coagulated. 

They  were  isolated  from  coagu- 
lated milk. 

Streptococcus  albus  (Tils).— 
Cocci  forming  motile  chains. 

Colonies  are  flat  and  circular, 
with  white  periphery  and  dark 
nucleus,  rapidly  liquefying. 

Inoculated  in  the  depth  of  gela- 
tine there  is  rapid  liquefaction  in 
the  track  of  the  needle,  and  a 
white  deposit. 

On  potato  they  form  a  white 
slimy  layer. 

They  were  found  in  water. 

Streptococcus  bombycis  (p.  472). 

Streptococcus  brevis  (Lingels- 
heim). — Cocci  singly,  in  pairs 
and  chains,  of  eight  to  ten 
elements. 

Colonies  on  gelatine  are  circular 
and  very  minute. 

Inoculated  in  the  depth  of  gela- 
tine there  is  a  funnel-shaped  cavity 
near  the  surface,  and  below  this, 
in  the  track  of  the  needle,  small 
isolated  colonies. 

On  agar  a  yellowish-grey  film 
develops  along  the  line  of  inocula- 
tion. 

On  potato  there  is  a  copious  white 
growth  in  forty-eight  hours. 

Broth  is  made  turbid. 

They  were  isolated  from  healthy 
saliva. 


570 


DESCRIPTION   OF   SPECIES. 


Streptococcus  cadaveris  (Stern- 
berg).— The  description  corresponds 
with  that  of  Streptococcus  pyo- 
genes. 

Inoculated  in  the  depth  of  gela- 
tine the  colonies  are  said  to  be 
larger  and  more  opaque. 

On  the  surface  of  agar  they  form 
a  thin  translucent  layer. 

In  broth  little  flocculi  develop, 
composed  of  chains  in  which  in 
some  cases  the  elements  varied 
considerably  in  size. 

They  were  obtained  from  the 
liver  in  a  fatal  case  of  yellow  fever. 

Streptococcus  coli  gracilis 
(Escherich). — Cocci  from  -2  to  '4  p, 
in  diam.,  forming  chains  composed 
of  from  six  to  twenty  elements. 
Some  elements  in  a  chain  are  irre- 
gular in  form,  and  show  transverse 
fission. 

The  colonies  are  spherical  and 
sink  down  in  the  liquefied  gela- 
tine. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  in  the  track 
of  the  needle  on  the  second  day, 
and  a  white  deposit  forms  at  the 
bottom  of  the  liquid.  In  about  a 
week  the  gelatine  is  completely 
liquefied. 

On  agar  there  is  a  very  slight 
growth. 

On  blood-serum  small  scales 
develop. 

On  potato  the  growth  is  com- 
posed of  small  white  prominent 
colonies. 

Milk  is  coagulated. 

They  were  found  in  the  evacua- 
tions of  healthy  infants. 

Streptococcus  conglomeratus 
(Kurth). — Cocci  and  chains,  identi- 
cal with  Streptococcus  pyogenes. 

They  form  an  adherent  film  at 
the  bottom  of  the  tube,  which  is 
not  broken  up  by  agitation.  This 
is  observed  in  other  varieties  of 
Streptococcus  pyogenes,  and  is  not 
sufficient  to  distinguish  it. 

They  are  pathogenic  in  mice. 

They  were  isolated  from  cases  of 
scarlet  fever. 

Streptococcus  flavus  desidens 
(Fliigge).— Cocci,  diplococci,  and 
short  chains.  They  form  yellowish- 


white  colonies,  which  gradually 
sink  down  in  the  gelatine. 

Inoculated  in  the  depth  of  gela- 
tine the  cocci  form  china-white, 
confluent  masses  in  the  track  of 
the  needle,  and  on  the  surface  a 
yellowish-brown  slimy  layer. 

They  occur  in  air  and  in  water, 
and  were  originally  isolated  from 
contaminated  cultures. 

Streptococcus  giganteus 
urethras  (Lustgarten).— Cocci  -8 
to  1  p.  in  diam.,  forming  chains 
composed  of  several  hundred 
elements.  In  description  they 
correspond  with  Streptococcus 
pyogenes. 

They  do  not  grow  at  the  tem- 
perature of  the  room. 

Colonies  on  agar  are  transparent 
and  iridescent. 

They  were  isolated  from  the 
healthy  urethra. 

Streptococcus  Havaniensis 
(Sternberg). — Cocci  '6  to  -9  /u  in 
diam.,  forming  long  chains,  com- 
posed of  cocci,  in  pairs,  and  oval 
elements  showing  transverse  divi- 
sion. 

This  streptococcus  is  probably 
a  variety  of  Streptococcus  pyo- 
genes. 

They  were  found  in  the  acid 
vomit  of  a  yellow-fever  patient. 

Streptococcus  in  contagious 
mammitis  in  cows  (Nocard  and 
Mollereau). — Cocci  spherical  or 
oval,  united  in  long  chains. 

Colonies  are  spherical,  granular, 
pale-yellow,  or  brownish  by  trans- 
mitted light. 

The  cocci  inoculated  in  the  depth 
of  gelatine  produce  a  granular 
filament  in  the  track  of  the 
needle. 

On  the  surface  of  nutrient  gela- 
tine minute  spherical  colonies  are 
formed,  which  are  bluish  by  re- 
flected light. 

Injected  into  the  mammary  gland 
of  cows  and  goats  they  produce 
mastitis. 

They  were  isolated  from  the  niilk 
of  cows  suffering  from  contagious 
mammitis. 

From  the  description  this  strepto- 
coccus appears  to  be  closely  related 


DESCRIPTION   OF    SPECIES. 


571 


to,  if  not  identical  with,  Strepto- 
coccus pyogenes  bovis(Crookshank). 
Streptococcus  in  progressive 
tissue  necrosis  in  mice.— Koch 
produced  a  disease  in  mice  by  sub- 
cutaneous injection  of  putrid  blood. 
In  tissue  sections  a  chain  coccus 
was  found  which  was  similar  to 
Streptococcus  pyogenes. 


FIG."  229.— STREPTOCOCCUS  IN  PRO- 
GRESSIVE TISSUE  NECROSIS  IN  MICE. 
a,  Necrotic  cartilage  cells,  and  (b) 
chains  in  masses ;  c,  isolated  chains. 
(Koch.) 

Streptococcus     in     Strangles 

(Schutz). — Cocci  forming  long 
chains,  which,  it  is  said,  do  not  grow 
on  nutrient  gelatine  or  agar,  but 
form  a  transparent  iridescent  cul- 
ture on  blood  serum.  Cultures  in 
broth  produced  the  disease  in  horses 
and  mice.  Rabbits,  guinea-pigs, 
and  pigeons  are  not  affected. 

Strangles  is  a  disease  of  the  horse, 
associated  with  suppuration  of  the 
glands  of  the  head  and  neck,  prin- 
cipally in  the  sub-maxillary,  sub- 
parotideal,  and  retro-pharyngeal 
regions.  Schutz  found  that  the  pus 
contains  streptococci  and  produces 
a  fatal  disease  in  mice. 

Streptococcus  liquefaciens 
(Sternberg). — Spherical  and  oval 


cocci,  *4  to  '6  /z  in  diam.,  singly,  in 
pairs  and  short  chains. 

Inoculated  in  the  depth  of  gela- 
tine liquefaction  occurs  rapidly  in 
the  track  of  the  needle,  and  in  a 
week  the  gelatine  is  completely 
liquefied,  slightly  opalescent,  and  a 
scanty  deposit  forms  at  the  bottom 
of  the  tube. 

In  the  depth  of  agar  a  filament 
is  formed  composed  of  closely- 
crowded  colonies. 

On  potato  a  thin  and  limited 
dry  white  layer  is  formed  along  the 
line  of  inoculation  in  four  to  five 
days. 

They  are  non-pathogenic. 

They  were  isolated  from  the  liver 
and  intestines  of  fatal  cases  of 
yellow  fever. 

Streptococcus  mirabilis  (Ros- 
coe  and  Lunt). — Cocci  4  p  in  diam., 
singly,  and  in  long  chains. 

The  growth  on  nutrient  media  is 
very  scanty. 

In  broth  the  growth  is  composed 
of  a  mass  of  delicate  filaments 
which  collect  at  the  bottom  of  the 
liquid. 

They  were  isolated  from  sewage. 

Streptococcus  of  Bonome.— 
Cocci  forming  chains.  They  corre- 
spond in  description  with  Strepto- 
coccus pyogenes,  but,  it  is  said, 
they  do  not  grow  in  gelatine  or 
on  blood-serum,  and  they  are  said 
to  be  distinguished  by  the  characters 
of  the  colonies  on  agar  plates. 

They  are  pathogenic. 

Inoculated  in  rabbits  and  white 
mice  they  produce  symptoms 
similar  to  those  produced  by  in- 
oculations of  the  pneumococcus. 
Sub-cultures  rapidly  lose  their 
virulence. 

They  were  isolated  from  cases 
of  cerebro-spinal  meningitis. 

Streptococcus  of  Manneberg  — 
Cocci  -9  \i  in  diam.,  singly,  in  pairs, 
and  in  chains. 

Inoculated  in  the  depth  of  gela- 
tine a  white  filament  forms  along 
the  track  of  the  needle  composed 
of  minute  colonies.  In  about  a 
month  the  filament  is  replaced  by 
a  funnel  of  semi-liquefied  gelatine. 

On    the    surface    of    agar    the 


572 


DESCRIPTION   OF   SPECIES. 


growth  resembles  Streptococcus 
pyogenes. 

"On   potato  they    form    a   slimy 
layer. 

Milk  is  rapidly  coagulated. 

They  are  pyogenic  in  dogs  and 
rabbits.  Injected  into  the  veins 
they  produce  inflammation  of  the 
kidneys. 

They  were  isolated  from  the 
urine  in  a  case  of  Bright' s  disease. 

Streptococcus  perniciosus 

psittacorum  {Parrot  disease). — 
Cocci,  singly,  in  chains,  and  in 
zooglcea,  have  been  described  in 
connection  with  a  disease  of  the 
grey  parrot  (Psittacus  erithacus). 
This  disease  is  fatal  to  about  80  per 
cent,  of  these  parrots  imported  to 
Europe.  They  suffer  from  diarrhoea 
and  general  weakness  ;  their  feathers 
are  ruffled ;  their  wings  hang 
loosely,  and  their  eyelids  close  ; 
convulsions  set  in,  and  death  fol- 
lows. At  the  autopsy  greyish 
nodules  are  found  in  the  lungs,  liver, 
spleen  and  kidney.  In  and  around 
the  capillaries  of  these  nodules,  and 
in  the  blood  of  the  heart,  the  cocci 
are  found  in  great  numbers  in 
zooglcea,  and  more  rarely  in  chains. 
Inflammatory  change  in  the  sur- 
rounding tissue  is  absent. 

Streptococcus  pyogenes  (p.  178). 

Streptococcus  radiatus 
(Fliigge). — Cocci  less  than  1  ^  in 
diam.,  singly,  in  small  masses,  and 
sometimes  in  short  chains. 

Colonies  appear  in  twenty-four 
hours.  They  are  white,  with  a 
yellowish-green  sheen  ;  later  they 
liquefy  the  gelatine  and  develop  a 
circlet  of  rays. 

Inoculated  in  gelatine,  isolated 
centres  form  along  the  track  of  the 
needle  which  throw  out  horizontal 
rays.  At  the  same  time  a  funnel- 
shaped  area  of  liquefaction  forms 
very  slowly  in  the  upper  part. 

On  potato  the  growth  is  yel- 
lowish-brown. 

They  occur  in  air  and  in  water. 

Streptococcus  septicus  (Fliigge). 
— Cocci  in  chains,  indistinguishable 
microscopically  from  Streptococcus 
pyogenes. 

Colonies  on  gelatine  grow   more 


slowly  than  those  of  most  strepto- 
cocci. 

They  are  pathogenic.  Mice  die 
in  forty-eight  to  seventy-two  hours 
after  subcutaneous  inoculation  of 
a  minute  quantity  of  a  cultivation. 
During  the  last  twenty-four  hours 
there  is  a  distinct  motor  and  sensory 
paralysis  of  the  hind  legs.  In 
rabbits  inoculation  of  the  ear  pro- 
duces local  redness,  then  a  general 
disease,  and  death  in  two  or  three 
days. 

They  were  found  by  Nicolaier,and 
independently  by  Guarneri,  in  earth. 

Streptococcus  septicus  lique- 
faciens  (Babes).— Cocci  -3  to  -4  ^ 
in  pairs,  in  short  chains. 

Inoculated  in  the  depth  of  gela- 
tine a  granular  filament  forms  in 
twenty-four  hours  along  the  track 
of  the  needle,  followed  by  lique- 
faction of  the  gelatine  forming  a 
funnel  in  which  the  gelatine  is 
clouded  ;  flat,  whitish  deposits 
form  on  the  side  of  the  funnel. 

On  the  surface  of  agar  minute 
shining,  transparent  colonies  are 
formed. 

On  blood-serum  the  growth  is 
almost  invisible. 

They  are  pathogenic.  Subcu- 
taneous injection  in  mice  and 
rabbits  produces  local  inflammation 
and  oedema,  followed  by  death  in 
about  a  week. 

They  were  found  in  the  blood 
and  organs  of  a  child  which  had 
died  of  septicaemia  complicating 
scarlet  fever. 

Streptococcus  yermiformis 
(Tils). — Cocci  forming  motile 
chains. 

Colonies  are  yellowish-white,  the 
central  portion  finely  granular, 
the  periphery  radiated. 

Inoculated  in  the  depth  of  gela- 
tine there  is  rapid  liquefaction, 
and  a  yellowish  deposit  at  the 
bottom  of  the  liquid. 

On  potato  the  culture  forms  a 
dirty-yellow  layer. 

They  were  found  in  water. 

Streptothrix  actinomycotica 
(p.  431). 

Streptothrix  alba  (Gasparini). 
— A  variety  of  Actinomyces  bovis. 


DESCRIPTION   OF   SPECIES. 


573 


Streptothrix      asteroides 

{ Ooxpora  asteroides,  Sauvageau 
and  Radais  ;  Cladothrix  asteroides, 
Eppinger).  —  Branching  filaments 
which  form  on  the  surface  of 
grape-sugar-agar  a  whitish  growth, 
which  is  later  of  a  brownish-yellow 
colour. 

Broth  remains  clear,  and  small 
pellicles  float  on  the  surface  re- 
sembling drops  of  stearin. 

On  potato  they  form  snow-white 
points,  which  turn  brick-red  in 
colour,  and  are  later  covered  with 
a  delicate  white  efflorescence. 

The  streptothrix  is  pathogenic 
in  rabbits  and  guinea-pigs. 

It  was  isolated  from  pus. 

Streptothrix  aurantiaca 
(Oospora  aurantiaca,  Sauvageau  and 
Radais,  and  Doria). — Similar  to 
Streptothrix  asteroides. 

Streptothrix  carnea  (Doria; 
Oospora  carnea,  Sauvageau  and 
Radais). — Similar  to  Streptothrix 
asteroides,  but  the  cultures  on 
gelatine  are  pink. 

They  are  not  pathogenic. 

Streptothrix  chromogenes 
(Gasparini  ;  Oospora  chromogenes, 
Lehmann  and  Neumann). — Culti- 
vated on  the  surface  of  gelatine  the 
filaments  produce  a  chalky  growth, 
and  the  jelly  is  coloured  brown,  and 
is  slowly  liquefied. 

i  On  potato  the  growth  is  yellowish 
or  brown,  and  the  potato  itself  is 
coloured  dark  brown  or  black. 

The  streptothrix  has  been  isolated 
from  air  and  water  and  the  con- 
tents of  the  stomach. 

Streptothrix  farcinica  (Bacille 
du  farcin  de  bceuf,  Nocard  ;  Oospora 
facinica,  Sauvageau  and  Radais). 

Inoculated  on  the  surface  of 
gelatine  there  is  in  about  two  weeks 
a  very  scanty  granular  growth. 

In  broth  greyish  pellicles  develop 
with  a  dusty  surface.  They  are 
pathogenic  in  cattle,  guinea-pigs, 
and  sheep. 

They  were  isolated  from  the 
disease  known  as  farcin  de  bceuf. 

Streptothrix  Forsteri  (Cohn). 
— Cocci  rods,  and  leptothrix  threads. 
The  threads  are  twisted  in  irregular 
spirals,  and  branch  sparingly  and 


irregularly.  Screw-forms  are  pro- 
duced by  the  threads  breaking  up 
into  small  pieces. 

Colonies  slowly  liquefy  gelatine. 
On    agar  they    form   a    whitish 
growth. 

In  broth  they  form  shining 
masses,  floating  in  clear  liquid. 

They  occur  in  the  lachrymal 
canals  of  the  human  eye,  in  the 
form  of  closely  felted  masses,  and 
in  the  air,  and  in  fresh-  and  sea- 
water. 

Streptothrix  Hofmanni  (Micro- 
myces  Hoffmanni,  Gruber  ;  Oospora 
Hoffmanni,  Sauvageau  and  Radais). 
—The  filaments  flourish  in  the  ordin- 
ary culture  media  with  the  addition 
of  sugar,  but  they  do  not  grow  on 
potato. 

They  produce  suppuration  in 
rabbits. 

They  were  isolated  from  the  air. 
Streptothrix  liquefaciens 
(Cladothrix  liquefaciens,  Garten). — 
A  variety  of  Actinomyces  bovis. 
Streptothrix  madurae  (p.  449). 
Streptothrix  musculorum 
SUis   (Actinomyces    suis,   Bunker). 
— A  variety  of  actinomyces  found 
in  the  muscles  of  swine. 

Streptothrix  odorif era  (Oospora 
odor  if  era,  Rullmann).  Probably 
identical  with  Oospora  chromogenes. 
Streptothrix  violacea  (Oospora 
violacea,  Sauvageau  and  Radais, 
and  Doria. — This  streptothrix 
liquefies  gelatine,  and  gives  it  a 
pale  wine-red  colour. 

Agar  is  coloured  a  violet  tint, 
and  potato  becomes  a  reddish- 
brown. 

Urobacillus  Dnclauxi 
(Miquel).— Rods  '6  to  -8  p  in 
diam.,  and  filaments  2  to  10  /*  in 
length.  Motile.  Spore-formation 
present. 

In  gelatine  containing  ammonia 
or  urea  they  develop  in  the  track 
of  the  needle  and  cause  liquefac- 
tion. The  liquefied  gelatine  is 
viscid. 

Broth  containing  ammonia  be- 
comes turbid,  a  sediment  forms,  and 
the  liquid  gives  off  an  unpleasant 
odour. 

They  occur  in  sewage. 


574 


DESCRIPTION   OF   SPECIES. 


Urobacillus  Freudenreichi 
(Miquel). — Rods  1  to  1-3  p  in  width, 
and  filaments  5  to  6  n  in  length. 

Colonies  circular,  white. 

Inoculated  in  the  depth  of  gela- 
tine growth  occurs  in  the  track  of 
the  needle,  and  a  pure  white  growth 
on  the  surface,  followed  by  slow 
liquefaction. 

In  broth  they  produce  turbidity. 

They  decompose  urea. 

They  occur  in  air,  sewage  and 
dust. 

Urobacillus  Maddoxi  (Miquel). 
—Rods  1  p.  in  width,  3  to  6  p,  in 
length,  and  involution  forms. 

Inoculated  in  the  depth  of  gela- 
tine containing  urea  they  produce 
white  colonies  and  crystals. 

In  broth  they  produce  turbidity. 

They  decompose  urea. 

They  occur  in  sewage. 

Urobacillus  Pasteuri  (Miquel). 
— Rods  attaining  1-2  /A  in  width, 
and  4  to  6  /u.  in  length,  singly  and 
in  pairs.  Spore-formation  present. 

They  grow  in  ammoniacal  gela- 
tine, slowly  liquefying  it  and  form- 
ing crystals.  The  liquefied  gelatine 
is  viscid. 

They  ferment  urine,  producing 
a  copious  deposit  of  crystals. 

They  were  isolated  from  decom- 
posing urine. 

Urobacillus  Schutzenbergi 
(Miquel). — Short  rods  '5  p.  in  width, 
1  n  in  length. 

They  rapidly  liquefy  gelatine. 

On  agar  they  form  a  white  layer. 

They  grow  readily  in  broth, 
especially  after  the  addition  of 
urea.  The  liquid  is  made  cloudy, 
but  after  a  few  days  it  becomes 
clear  again. 

They  occur  in  water. 

Vibrio  rugula  (Miiller).— Rods 
and  threads,  6  to  16  p  long,  about 
•5  to  2-5  \i  thick.  The  rods  are 
either  simply  bowed,  or  possessed 
of  one  shallow  spiral  (Fig.  230). 


They  bear  a  flagellum  at  each  end. 
The  rods  form  swarms  when  caus- 
ing decomposition,  and  then,  or 
after,  grow  out  into  threads,  curved 
in  a  screw-like  manner.  In  the 


FIG.  230.— VIBRIO  RUGULA,  x  1020. 
A.  Bowed  threads.  B.  Slightly- 
curved  rods.  C.  Rods  swollen  pre- 
paratory to  spore-formation.  D. 
Rods  swollen  at  the  spore -forming 
end.  E.  Various  stages  of  the 
developing  spores.  (Prazmowski. ) 


next  stage  of  development  the  rods 
cease  to  move,  and  become  swollen 
with  granular  contents.  One  ex- 
tremity develops  an  enlargement, 
giving  the  rod  the  appearance  of 
a  pin.  The  spore  formed  by 
the  contraction  of  the  plasma  in 
the  swollen  end  finally  becomes 
globular. 

The  vibrios  appear  in  vegetable 
infusions,  causing  fermentation  of 
cellulose. 


APPENDICES 


575 


APPENDIX  I. 

YEASTS  AND   MOULDS. 

Yeast-fungi  and  mould-fungi,  like  bacteria  or  fission-fungi,  are- 
achlorophyllous  Thallophytes.  They  belong  to  two  separate  orders — 
the  Saccharomycetes  and  Hyphomycetes — which  are  intimately  related 
to  each  other,  but  quite  distinct  from  bacteria.  Their  germs  occur 
widely  distributed  in  air,  soil  and  water,  and  are  constantly 
encountered  in  bacteriological  investigations.  In  addition,  many 
species  are  of  hygienic  and  pathological  interest  and  importance  in 
being  either  accidentally  associated  with,  or  the  cause  of  various 
morbid  processes  and  fermentations.  For  a  complete  account  of 
all  the  described  species  and  full  details  of  the  various  forms  of 
development,  reference  must  be  made  to  botanical  and  other 
works.*  A  description  of  certain  species  is  appended  here,  and  may 
afford  some  useful  information  to  the  worker  in  a  bacteriological 
laboratory. 

YEAST- FUNGI  OR  SACCHAROMYCETES. 

Saccharomyces  cerevisise  (Torula  cerevisice). — Cells  round  or 
oval,  8  to  9  //,  long,  singly  or  united  in  small  chains.  Spores 
occur  three  or  four  together  in  a  mother-cell,  4  to  5  /A  in  diam. 
S.  cerevisise,  S.  pastorianus  and  S.  ellipsoideus  are  active  alcoholic 
ferments.  According  to  Jorgensen  they  will  produce  in  fourteen 
days  in  beer-wort  from  4  to  6  per  cent.,  by  volume,  of  alcohol. 

Saccharomyces  ellipsoideus  (Hansen).  I. — Elliptical  cells, 
mostly  6  /A  long,  singly  or  united  in  little  branching  chains.  Two  to 
four  spores  found  in  a  mother-cell,  3  to  3*5  //,  in  diam.  Cultivated 
on  the  surface  of  wort-gelatine  they  produce  in  eleven  to  fourteen 
days,  at  25°  C.,  a  net-like  growth  by  which  they  can  be  recognised 

*  Sachs,  Text-book  of  Botany  ;  Jorgensen,  Micro-organisms  and  Fermen- 
tation. 

577  37 


578  APPENDICES. 

with  the  naked  eye.  II.— Round,  oval,  and  rarely  elongated 
cells.  They  produce  yeast-turbidity.  There  are  two  so-called 
disease-yeasts  allied  to  this  species.  The  colonies  of  one  kind  form 
a  network.  This  yeast  causes  turbidity  in  beer,  and  a  bitter  after- 
taste. In  the  other  kind  the  colonies  are  sharply  denned.  It 
produces  a  disagreeable  aromatic  taste  to  beer,  and  an  astringent 
after- taste.  It  is  widely  distributed,  and  is  the  principal  agent  in 
.accidental  fermentation. 

Saccharomyces  eonglomeratus  (Reess). — Cells  round,  5  to 
6  fji  in  diam.,  united  in  clusters,  consisting  of  numerous  cells 
produced  by  budding  from  one  or  a  few  mother- cells.  There  are 
2  to  4  spores  in  each  mother-cell.  They  occur  on  rotting  grapes 
and  in  wine  at  the  commencement  of  fermentation. 

Saccharomyces  exiguus  (Reess). — Conical  or  top-shaped 
cells,  5  p  long,  and  reaching  2'5  /x,  in  thickness,  in  slightly 
branching  colonies.  Spore-forming  cells  are  isolated,  each  contain- 
ing 2  or  3  spores  in  a  row.  They  occur  in  the  after-fermentation 
of  beer;  but,  according  to  Hansen,  they  do  not  produce  disease 
in  beer. 

Saccharomyces  Jb'rgensenii  (Lasche). — Cells  small,  round 
or  oval.  On  the  surface  of  wort-gelatine  the  culture  is  greyish- 
white,  and  the  gelatine  is  slowly  liquefied.  They  ferment  saccharose 
and  dextrose,  but  not  maltose.  When  grown  in  wort  with  other 
yeasts  they  are  rapidly  crowded  out. 

Saccharomyces  pastorianus,  I. — Cells  oval  or  club-shaped. 
Colonies  consist  of  primary  club-shaped  links,  18  to  22  p  long, 
which  build  lateral,  secondary,  round  or  oval  daughter-cells,  5  to 
6  p  long.  Spores  2  to  4.  They  occur  in  the  after- ferment  at  ion 
of  wine,  fruit-wines,  or  fermenting  beer,  and  in  the  air  of  breweries. 
They  produce  a  bitter  taste  and  unpleasant  odour  and  turbidity 
in  beer.  II. — Cells  mostly  elongated,  but  also  oval  or  round. 
Cultivated  on  the  surface  of  gelatine  and  yeast- water  a  growth  is 
produced  with  smooth  edges,  by  which  it  can  be  differentiated 
from  No.  III.  They  occur  in  the  air  of  breweries,  but  do  not 
produce  disease  in  beer.  III. — They  produce  yeast-turbidity  in 
beer.  On  the  surface  of  yeast- water  gelatine  the  cultures,  after 
sixteen  days,  have  hairy  edges. 

Saccharomyces  apiculatus. — Cells  lemon -shaped,  both  ends 
bluntly  pointed,  6  to  8  /x  long,  2  to  3  p  wide.  Budding  occurs  only 
at  the  pointed  ends.  Rarely  united  in  colonies.  Spores  unknown. 
They  occur  with  other  yeasts  in  various  accidental  fermentations 
-and  in  ripe  fruits. 


YEASTS   AND   MOULDS.  579 

Saccharomyces  sphsericus. — Cells  varying  in  form ;  the 
basal  ones  of  a  colony  oblong  or  cylindrical,  10  to  15  /x  long, 
5  /x  thick ;  the  others,  round,  5  to  6  /A  in  diam.  United  in  ramified 
families.  Spores  unknown. 

Saccharomyces  anomalus  (Hansen). — Cells  small,  oval,  and 
sometimes  elongated.  Spores  are  hemispherical,  with  projecting  rims 
at  the  base.  They  were  found  in  impure  brewery  yeast. 

Saccharomyces  mycoderma  (Mycoderma,  cerevisice  et  vini). — 
Cells  oval,  elliptical,  or  cylindrical,  6  to  7  /x  long,  2  to  3  /x  thick, 
united  in  richly-branching  chains.  Spore-forming  cells  may  be 
20  /x  long.  Spores  1  to  4  in  each  mother-cell.  The  colonies  in 
gelatine  are  greyish  and  filmy.  They  form  the  so-called  "  mould  " 
on  fermented  liquids,  arid  develop  on  the  surface  without  exciting 
fermentation.  When  forced  to  grow  submerged,  a  little  alcohol  is 
produced,  but  the  fungus  soon  dies.  They  occur  on  wine,  beer,  fruit- 
juices  and  sauerkraut. 

Saccharomyces  albicans  (Oidium  albicans,  Fungus  of  thrush). 
— Cells  round,  oval,  or  cylindrical,  3*5  to  5  /x  thick  ;  the  cylindrical 
cells  10  to  20  times  as  long  as  they  are  thick.  The  bud- colonies 
mostly  consist  of  rows  of  cylindrical  cells,  from  the  ends  of  which 
oval  or  round  cells  shoot  out.  Spores  form  singly  in  roundish  cells. 
In  plate-cultivations  the  colonies  are  pure  white.  In  the  depth 
of  gelatine  a  filament  is  formed  composed  of  white  colonies,  some 
with  ray-like  processes  extending  into  the  gelatine.  On  potato  the 
fungus  forms  a  rapid  white  growth,  and  on  bread  also.  They 
can  be  easily  cultivated  in  a  nutrient  solution  containing  sugar 
and  ammonic  tartrate.  The  cells  germinate  according  to  the  rich- 
ness of  the  fluid  in  sugar ;  they  either  grow  into  long  threads, 
or,  in  a  very  strongly  saccharine  solution,  many  daughter- cells  are 
formed  and  bud  out  in  various  directions.  According  to  Klemperer 
the  thrush-fungus  is  pathogenic  in  rabbits,  death  taking  place 
twenty-four  to  forty-eight  hours  after  an  intravenous  injection  of 
a  pure- culture.  Long  mycelial  threads  are  found  in  the  internal 
organs.  They  occur  on  the  mucous  membrane  of  the  mouth, 
especially  of  infants,  in  greyish -white  patches,  which  consist  of 
epithelium,  bacteria,  yeasts,  and  the  mycelia  of  various  moulds. 

Saccharomyces  pyriformis  (Marshall  Ward). — Cells  oval. 
They  convert  saccharine  solutions  containing  ginger  into  ginger- 
beer.  They  occur  with  other  micro-organisms  in  the  so-called 
"  ginger-beer  plant." 

Saccharomyces  glutinis. — Cells  round,  oval,  or  short 
cylinders,  5  to  11  /x  long,  4  /x  wide,  isolated,  or  united  in  twos. 


580  APPENDICES. 

Cell- membrane  and  contents  are  colourless  in  the  fresh  state,  but 
when  dried  and  re-moistened  possess  a  pale-reddish  nucleus  in  the 
middle.  Spore-formation  unknown.  They  form  rose-coloured,  slimy 
spots  on  starch  paste,  and  on  sterilised  potatoes.  The  colouring 
matter  is  not  changed  by  acids  or  alkalies. 

Saccharomyces  ilieis  (Gronlund).— Cells  spherical.  Spore- 
formation  present  without  vacuoles.  Cultures  on  the  surface  of 
gelatine  have  a  powdery  appearance.  They  produce  about  2 '8 
per-  cent.,. by  volume,  of  alcohol  in  beer-wort,  and  cause  a  disagree- 
able, bitter  taste.  They  were  obtained  from  the  fruit  of  Ilex 
aquifolium. 

Saccharomyces  aquifolii  (Gronlund). — Cells  large  and 
spherical.  Spores  contain  vacuoles.  Cultures  on  gelatine  are 
variable,  smooth  and  shining,  or  powdery.  They  produce  about 
3-7  per  cent,  alcohol  in  beer- wort,  and  cause  a  sweet  taste  with 
bitter  after-taste.  They  also  were  obtained  from  the  fruit  of  Ilex 
aquifolium. 

Saccharomyces  Marxianus  (Hansen). — Cells  elongated. 
They  develop  a  mycelial  growth  on  solid  nutrient  media.  They 
occur  on  grapes. 

Saccharomyces  membransefaeiens  (Hansen). — Cells  elon- 
gated and  vacuolated.  Spore-formation  abundant.  Cultivated  on 
wort-gelatine  they  produce  circular,  flattened  and  wrinkled  colonies, 
greyish,  and  sometimes  with  a  reddish  tinge.  The  gelatine  is  slowly 
liquefied.  They  occur  in  the  slimy  secretion  of  the  roots  of  the 
elm,  and  were  also  isolated  from  well-water. 

Saccharomyces  Hansenii  (Zopf). — Cells  with  small  spherical 
spores.  They  set  up  alcoholic  fermentation  in  solutions  containing 
sugar.  They  were  found  in  cotton- seed  flour. 

Saccharomyces  Ludwigii. — Cells  irregular  in  form,  oval, 
bottle-shaped,  lemon-shaped,  and  elongated,  and  mycelial  filaments. 
On  wort- gelatine  the  growth  is  greyish  or  yellowish. 

Saccharomyces  acidi  lactici  (Grotenfelt). — Cells  oval,  2  to 
4-35  /A  in  length,  and  1'5  to  2'9  /x  in  width.  Colonies  on  nutrient 
gelatine  are  porcelain-white.  They  coagulate  milk. 

Saccharomyces  minor  (Engel). — Cells  spherical.  Spore- 
formation  present.  They  are  said  to  be  the  most  active  ferment 
in  the  fermentation  of  bread. 

Saccharomyces  rosaceus  (Pink  Torula). — Cells  9  to  10  /x 
in  diam.  They  form  a  coral- pink  growth  in  nutrient  gelatine, 
nutrient  agar-agar,  or  on  sterilised  potatoes.  They  are  present 
in  the  air. 


YEASTS   AND   MOULDS.  581 

Saccharomyces  niger  (Black  Torula).— Cells  also  present 
in  the  air.  Cultivated  in  nutrient  gelatine  they  form  a  black 
crust  (Fig.  231). 

MOULD-FUNGI  OR  HYPHOMYCETES. 

The  mould-fungi  have  been  divided  into 
five  orders  :  Hypodermii,  Phycomycetes,  Asco- 
mycetes,  Basidiomycetes  and  Myxomycetes. 
The  following  species,  with  the  orders  to 
which  they  belong,  are  of  especial  interest : — 

HYPODERMII. 

Ustilago  carbo  (Mildew,  Smut). — Spores 
brown,  circular  ;  episporium  smooth  ;  sporidia, 
ovoid  cells.  The  spores  or  conidia  occur  as 
a  black  powder  in  the  ears  and  panicles  of 
wheat,  barley  and  oats. 

Tilletia    caries. — Spores    round,    pale 
brown ;  episporium  with  reticulated  thicken- 
ings.     In   germinating,   the    sporidia    grow 
out  radially  from  the  end  of   the  promyce- 
lium  ;  these,  at  their  lower  part,  conjugate     p^   231  _BLACR    TQR. 
by  a  cross  branch    and   separate  from   the         ULA>     pURE   CULTIVA- 
promycelium,  and  at  some  point  of  the  pair         TION  ON  POTATO. 
a   hypha    grows    out,    on   which    abundant 

secondary  sporidia  develop.  The  latter  are  long,  oval  cells,  which 
can  in  turn  germinate.  The  fungus  occurs  in  the  form  of  a 
stinking  powder  in  grains  of  wheat,  which  renders  the  meal  im- 
pure, and  gives  it  a  disagreeable  smell. 

Urocystis  occulta. — The  spores  consist  of  several  cells  united 
together;  partly,  large  dark-brown  cells  in  the  interior,  and  out- 
side, several  flat,  semicircular,  colourless  cells.  The  promycelium 
germinates  as  in  Tilletia,  but  the  cylindrical  cells  produce  a  hypha, 
without,  as  a  rule,  previous  conjugation.  They  occur  as  a  black 
powder  in  rye-straw  in  long  disintegrated  stripes,  which  are  at  first 
greyish.  The  affected  plant  produces  abortive  ears. 

Empusa  muscse. — A  spore  or  conidium  of  this  fungus 
alighting  upon  the  white  area  of  the  under  surface  of  the  body 
of  the  house-fly  germinates  into  a  hypha.  The  latter,  penetrating 
the  skin,  forms  toruloid  cells,  which  multiply  by  germination,  and 
are  disseminated  in  the  blood  throughout  the  body  of  the  fly. 


582  APPENDICES. 

These  cells  again  grow  into  hyphse,  which  penetrate  the  skin,  each 
forming  a  conidium,  which  is  cast  off  with  considerable  force.  The 
parasite  is  fatal  to  flies,  especially  in  the  autumn.  They  are  often 
observed  attached  to  the  walls  or  window-panes,  surrounded  by  a 
powdery  substance,  consisting  of  the  extruded  conidia. 

Empusa  radicans. — The  spores  form  long  hyphse,  which  pierce 
the  transparent  skin  of  the  caterpillar  of  the  cabbage  white  butter- 
fly. The  terminal  cells  ramify,  and  fill  the  body  of  the  caterpillar 
with  a  network  of  mycelial  filaments.  The  caterpillars  attacked 
become  restless,  then  motionless,  and  death  ensues. 

Tarichium  megaspernmm. — The  spores  are  black  in  colour, 
and  provided  with  a  thickened  episporium.  They  occur  at  the 
sides  and  ends  of  mycelial  threads,  attacking  caterpillars  (Agrotis 
segetum). 

PHYCOMYCETES. 

Saprolegnia. — Colourless  threads,  forming  dense  radiating  tufts, 
occur  on  living  and  dead  animal  and  vegetable  matter  in  fresh 
water.  The  filaments  penetrate  into  the  substratum,  and  branch 
more  or  less  in  the  surrounding  water.  The  cylindrical  ends  of  the 
threads  are  shut  off  by  a  septum — forming  zoosporangia,  or  mother- 
cells,  in  the  interior  of  which  a  number  of  spherical  zoospores 
develop.  These  are  set  free  through  an  apical  opening  in  the 
thread,  and  after  a  time  coming  to  rest,  give  rise  to  new  plants. 
In  the  sexual  mode  of  reproduction  a  spherical  bud,  the  oogonium, 
develops  at  the  end  of  a  mycelial  thread;  from  the  thread  small 
processes  or  antheridia  sprout  out  laterally  towards  the  oogonium 
and  blend  with  its  protoplasm.  The  latter  breaks  up  into  a  number 
of  oospores,  which  clothe  themselves  with  a  membrane  while  still 
within  the  mother- cell,  and,  eventually  being  set  free,  grow  into 
fresh  mycelial  filaments.  The  fungus  attacks  fish  and  tritons, 
and  produces  a  diseased  condition  of  the  skin,  which  may  be 
ultimately  fatal.  In  salmon  it  produces  the  common  "disease  of 
salmon." 

Peronospora  infestans. — The  conidia -bearers  of  this  fungus 
have  as  many  as  five  branches,  each  bearing  an  egg-shaped 
conidium.  The  contents  of  the  conidia  falling  off  and  reaching  a 
drop  of  moisture,  break  up  into  a  number  of  swarming  zoogonidia, 
which  in  turn  develop  upon  plants.  Fixing  themselves  to  the 
cuticle  of  the  host,  they  throw  a  germinating  filament  into  an 
epidermal  cell ;  after  piercing  first  its  outer  wall,  and  then  its  inner 


YEASTS   AND   MOULDS.  583- 

wall,  the  filament  reaches  an  intercellular  space,  where  the  mycelium 
develops.  This  continues  to  grow  and  spread  throughout  the  plant. 
In  tubers  it  can  hibernate  and  develop  in  the  young  shoots  in  the 
following  spring.  The  fungus  appears  in  the  form  of  brown 
patches  on  the  green  parts  of  the  plants,  especially  the  leaves. 
The  attacked  parts  wither  and  turn  yellow  or  brown  in  colour. 
If  the  under  surface  of  a  diseased  leaf  is  examined,  a  corresponding 
dark  spot  may  be  observed,  accompanied  with  a  faint  greyish-white 
bloom,  which  covers  it.  The  latter  consists  of  the  conidia- bearing 
branches. 

Pilobolus. — The  fruit  -hyphse  possess  spherical  receptacles 
containing  conidia.  When  ripe  the  receptacles  with  their  conidia 
are  detached  at  their  bases,  and  spring  by  their  elasticity  to  some 
distance.  The  fungus  occurs  as  glassy  tufts  on  the  excrement  of 
cows,  horses,  etc.  A  cultivation  can  generally  be  obtained  by 
keeping  fresh  horse-dung  under  a  bell-glass. 

Mucor  mucedo. — Hyphae  colourless,  simple  or  branched ;  spo- 
rangia yellowish-brown  or  black ;  spores  ovoid.  They  form  the 
familiar  white  mould  on  fruits,  bread,  potatoes  and  excreta,  and 
penetrate  into  the  interior  of  nuts  and  apples.  A  network  of 
fibrils  develops  in  the  substance  of  nutrient  gelatine,  with  forma- 
tion of  sporangia  on  the  free  surface.  The  germination  of  the 
spores  and  development  into  hyphse  can  be  observed  in  a  few 
hours  if  the  fungus  be  cultivated  in  a  decoction  of  horse-dung. 

Mucor  racemosus. — Hyphse  short ;  sporangia,  yellowish  to 
pale -brown ;  spores  round.  By  continued  cultivation  in  liquids 
saturated  with  carbonic  acid,  the  hyphse  become  still  shorter 
arid  exhibit  a  yeast-like  sprouting.  These  yeast-like  or  toruloid 
cells  can,  when  the  carbonic  acid  is  withdrawn,  germinate  into 
normal  mycelium.  They  occur  on  bread  and  decaying  vegetable 
matter. 

Mucor  stolonifer  (Lichtheiin). — Mycelium  grows  in  the  air  and 
then  bends  down  and  re-enters  the  nutrient  substratum  ;  sporangia 
black,  and  spores  globular.  The  mycelium  can  penetrate  through 
the  shell  of  eggs,  and  form  conidiophores  within  them. 

Mucor  aspergillus  (Lichtheim). — Fruit-hyphae  thinned  at  the 
base,  and  with  many  fork-like  divisions  ;  dark-brown  spores. 

Mucor  phycomyces  (Lichtheim). — Mycelium  thick-walled  ; 
olive-green  fruit-hyphse  ;  black  sporangia,  and  oblong  spores. 

Mucor  macroearpus  (Lichtheim). — Spindle-formed,  pointed 
spores. 

Mucor  fusiger  (Lichtheim). — Ovoid  spores. 


584  APPENDICES. 

Mucor  mellittophorus  (Lichtheim). — Spores  elliptical.  Found 
in  the  stomach  of  bees. 

Mucor  corymbifer  (Lichtheim). — This  fungus  forms  branched 
fruit-hyphae.  The  sporangia  have  a  smooth  membrane.  It  has 
been  found  in  the  external  auditory  meatus,  and  on  bread  it  forms 
a  dense  snow-white  growth.  Pathogenic  in  rabbits. 

Mucor  rhizopodiformis  (Lichtheim). — The  spores  of  Mucor 
rhizopodiformis  and  Mucor  corymbifer,  when  introduced  into  the 
vascular  system  of  rabbits,  can  germinate  in  the  tissues,  especially 
in  .the  kidneys,  where  they  set  up  haemorrhagic  inflammation. 
Dogs  are  immune,  and  only  artificial  mycosis  is  known.  It  occurs 
on  bread. 

Mucor  erectus. — Resembles  Mucor  racemosus.  It  occurs  on 
rotting  potatoes. 

Mucor  circinelloides. — Mycelium  much  branched,  and 
sporangium  carrier  is  curved. 

Mucor  spinosus. — Sporangia  chocolate.  Columella  has  short 
processes  or  spines. 

ASCOMYCETES. 

Oidium  Tuckeri. — Fruit-hyphae  bearing  single  ovoid  conidia. 
Observed  in  the  form  of  brown  patches,  covered  with  a  white  mildew- 
like  layer  on  the  leaves,  branches  and  young  fruit  of  the  vine, 
producing  "  grape  disease." 

Oidium  lactis. — Fruit-hyphae  simple,  erect  and  colourless, 
bearing  at  their  ends  a  series  or  chain  of  conidia.  In  some  cases, 
the  fruit-hypha  branches  beneath  the  chain  of  spores.  Spores  are 
short  cylinders.  The  conidia  germinate  into  filaments  of  varying 
length,  which  by  subdivision  form  septate  mycelial  hyphse ;  these 
and  their  branches  give  rise  in  turn  to  spores  or  conidia.  The 
fungus  is  deeply  stained  by  the  ordinary  aniline  dyes.  In  a  plate- 
cultivation  the  colonies  appear  as  white  points,  and  develop  into 
delicate  stellate  colonies  which  ultimately  coalesce  and  form  a  fine 
mycelial  network  covering  the  surface  of  the  gelatine.  The  gela- 
tine is  not  liquefied.  The  growth  on  the  surface  of  agar  is  similar 
to  that  on  gelatine.  The  fungus  occurs  in  sour  milk. 

Achorion  Schonleinii  (Fungus  of  favus).— Threads  branching 
at  right  angles.  Favus  in  man  forms  yellow  crusts  on  the  hairy  parts 
of  the  body.  The  crusts  are  composed  of  epidermis  and  mycelial 
filaments  and  spores.  In  plate-cultivations  whitish  colonies  are 
formed  surrounded  by  liquefied  gelatine.  Cultivated  on  the  surface 


YEASTS   AND   MOULDS. 


585 


of  gelatine  the  growth  resembles  that  of  Tricophyton  tonsurans,  but 
the  liquefaction  takes  place  more  slowly,  and  there  is  a  more  distinct 
yellow  colour.  On  agar  the  growth  is  white,  dry  and  firmly 
adherent. 

Tricophyton  tonsurans  (Fungus  of  ringworm).— Mycelial 
filaments  and  spores  occur  on  the  crusts  and  in  diseased  hairs. 
In  plate-cultivations  white  colonies  are  formed,  and  liquefaction 
quickly  follows.  In  test-tube  cultivations  the  gelatine  is  liquefied 


FIG.  232.— HEAD  AND  NECK  OF  CALF  WITH  ADVANCED  RINGWORM  (Broiori). 

and  the  fungus  forms  a  membrane  on  the  liquid  jelly  which  is  white 
above  and  yellow  beneath.  The  surface  of  the  growth  is  powdery 
In  man  the  disease  varies  in  appearance  in  different  parts  of  the 
body.  Cattle,  horses  and  dogs  also  suffer  from  ringworm  ;  but  sheep 
and  pigs  rarely,  if  ever.  The  disease  is  very  common  in  calves. 
Sometimes  a  small  portion  of  the  skin  is  diseased;  in  other  cases, 
the  head,  neck,  chest  and  abdomen,  or  even  the  whole  trunk,  may  be 
covered  with  scabs  or  crusts.  There  is  often  loss  of  hair  in  patches, 
and  the  skin  may  be  covered  with  scurf.  The  disease  is  transmissible 


586  APPENDICES. 

to  the  human  subject.  In  one  case,  according  to  Brown,  seven 
grooms  were  infected  on  the  arms  from  a  grey  pony  which  was 
suffering  from  the  disease  in  an  aggravated  form. 

Fungus  of  fowl-scab. — Fowls  are  liable  to  a  disease  similar 
to  favus.  According  to  Schiitz  this  disease  is  characterised  by 
greyish-white  patches  on  the  comb  and  wattles  of  fowls,  which  may 
extend  over  the  neck  and  body.  On  nutrient  gelatine  a  white 
mycelium  is  formed  ;  and  the  gelatine  is  liquefied,  and  acquires  a 
reddish  tint.  The  fungus  can  be  readily  cultivated  on  bread-paste, 
agar-agar  and  potato.  Cultures  inoculated  in  fowls  produce  the 
disease,  but  have  no  effect  on  mice  and  rabbits. 

Fungus  of  mouse-favus. — Mice  suffer  from  a  form  of  favus 
which  can  be  communicated  to  healthy  mice  by  inoculation  of  scabs 
or  infected  skin  (Nicolaier).  On  nutrient  agar  the  fungus  forms  a 
thick  mycelium,  at  first  white,  and  later  of  a  red  or  reddish-brown 
colour.  Mice  can  be  infected  with  cultures. 

Microsporon  furfur. — This  fungus  occurs  in  Pityriasis 
versicolor.  Grawitz  regarded  it  as  identical  with  Oidium  lactis,  and 
it  is  very  closely  related.  Cultivated  on  gelatine  the  jelly  is  hollowed 
out  and  the  mycelial  growth  sinks  down,  and  is  yellowish  in  colour. 

Oidium.  albicans. — Vide  Saccharomyces  albicans. 

Aspergillus  glaucus  (Eurotium  aspergillus  glaucus). — Mycelium 
at  first  whitish,  becoming  grey-green  or  yellow-green.  Spores 
grey-green,  thick -walled.  It  is  found  on  various  substances,  chiefly 
cooked  fruit,  and  is  non- pathogenic. 

Aspergillus  repens  (Eurotium  repens,  De  Bary). — Fruit-heads 
fewer  than  in  the  above,  which  are  at  first  pale  and  then  blue-green 
to  dark-green  in  colour.  Conidia  mostly  oval,  smooth,  colourless 
or  pale  to  grey-green. 

Aspergillus  flavus.— Gold-yellow,  greenish  and  brown  tufts. 
Fruit-heads  round,  yellow,  olive-green  or  brown.  Conidia  round, 
seldom  oval,  sulphur-yellow  to  brown  in  colour.  Saprophytic  in 
man,  pathogenic  in  rabbits. 

Aspergillus  fumigatus. — Greenish,  bluish  or  grey  tufts. 
Fruit-heads  long  and  conical.  Conidia  round,  and  rarely  oval, 
smooth,  mostly  pale  and  colourless,  This  fungus  occurs  on  bread,, 
and  has  been  found  in  the  human  lungs,  external  auditory 
meatus  and  middle  ear,  and  in  the  lungs  of  birds.  The  spores 
introduced  into  the  vascular  system  of  rabbits,  or  into  the  peritoneal 
cavity,  establish  metastatic  foci  in  the  kidneys,  liver,  intestines,, 
lungs,  muscles,  and  sometimes  in  the  spleen,  bones,  lymphatic 
glands,  nervous  system  and  skin. 


YEASTS   AND   MOULDS.  587 

Aspergillus  niger  (Eurotium  aspergillus  niger,  De  Bary).— 
Dark  chocolate-brown  tufts.  Conidia  round,  black-brown,  or  grey- 
brown  when  ripe.  This  mould  can  be  cultivated  readily  on  bread 
moistened  with  vinegar,  on  slices  of  lemon,  and  on  acid  fruits  and 
liquids.  It  nourishes  best  of  all,  according  to  Raulin,  in  a  liquid 
of  the  following  composition  : — 

Grammes. 

Water 1500' 

Sugar-candy      .......  70* 

Tartaric  acid      .......  4* 

Nitrate  of  ammonia  ......  4' 

Phosphate -6 

Carbonate  of  potassium     .....  '6 

„             ,,  magnesium   .....  '4 

Sulphate  of  ammonia         .         .         .         .         .  '25 

,,         ,,  zinc         ......  '07 

„  iron  -07 

Silicate  of  potassium           .....  '07 

It  was  also  found  that  the  fungus  grew  best  when  the  liquid 
was  spread  out  in  a  layer  2  or  3  cm.  in  depth  in  a  shallow  dish ; 
and  a  temperature  of  35°  C.  proved  to  be  the  most  favourable. 
The  abstraction  of  zinc  from  the, nutritive  liquid  reduced  the  weight 
of  a  crop  from  25  (the  average)  to  2  grammes,  and  the  presence 
of  T^OOOOO  Part  of  nitrate  of  silver,  or  ^-^  part  of  corrosive 
sublimate,  stopped  the  growth  altogether.  It  is  saprophytic  in  the 
living  body. 

METHOD  OF  EXAMINING  ASPERGILLUS  NIGER. 

Species  of  aspergillus  stain  intensely  with  carmine,  f  uchsine  or  methyl- 
violet  ;  but  to  examine  Aspergillus  niger  with  a  high  power  a  little 
special  technique  is  employed,  as  follows  : — A  drop  of  glycerine  is  placed 
on  a  clean  slide,  and  a  drop  of  alcohol  on  a  cover-glass.  With  a  fine  pair 
of  forceps  a  few  of  the  fruit-hyphae  with  their  black  heads  are  immersed 
in  the  alcohol.  The  cover-glass  is  then  turned  over  on  to  the  drop  of 
glycerine,  and  the  slide  held  in  the  flame  of  a  Bunsen  burner  till  the 
spores  or  conidia  are  dispersed.  To  make  a  permanent  preparation 
remove  the  cover-glass,  and  transfer  the  fruit-hyphae  so  treated  to  a 
mixture  of  glycerine  and  water  (1  to  5)  ;  a  drop  may  be  conveniently 
placed  ready  on  a  slide  provided  with  a  ring  of  Canada  balsam.  The 
specimen  is  then,  permanently  mounted  by  employing  a  circular  cover- 
glass,  and  surrounding  it  with  a  ring  of  cement  in  the  usual  way. 


588  APPENDICES. 

Aspergillus  ochraceus. — At  first  flesh-coloured,  and  then 
ochre-yellow  heads. 

Aspergillus  albus. — Pure-white  fruit-heads. 

Aspergillus  clavatus. — Club-shaped  fruit-heads  on  long  stems. 

Aspergillus  nidulans. — Bread  and  potatoes  acquire  a  reddish- 
brown  colour.  Pathogenic  in  rabbits.  Occurs  on  bread. 

Aspergillus  subfuscus. — The  growth  is  olive-yellow  in  colour. 
Pathogenic  in  rabbits.  Occurs  on  bread. 

Aspergillus  flavescens.  —  The  growth  is  yellowish-green. 
Pathogenic  in  dogs  and  rabbits.  Occurs  on  bread. 

Penicillium  glaucum. — Occurs  as  a  white,  and  later  a  blue 
green,  mould,  on  which  dew-like  drops  of  liquid  may  appear.  Its 
spores  are  present  in  large  numbers  in  the  air,  and  are  liable 
to  contaminate  cultivations.  The  fruit-hypha  bears  terminally  a 
number  of  branched  cylindrical  cells,  from  which  chains  of  greenish 
conidia  are  developed.  It  is  the  commonest  of  all  moulds. 

Botrytis  Bassiana — Hyphse  and  spores  colourless.  Hyphae 
usually  simple,  but  sometimes  united  in  arborescent  stems.  It  is 
the  cause  of  muscardine,  a  fatal  disease  of  silkworms,  and  occurs 
also  in  various  other  caterpillars  and  insects. 

Chionyphe  Carteri.— Mycelial  filaments  observed  by  Carter 
in  Madura  disease. 


APPENDIX    II. 


H^EMATOZOA. 

H^MATOZOA     IN     MAN,     BIRDS      AND      TURTLES. — H^MATOZOA      IN 
EQUINES,    CAMELS,    RATS   AND   FISH. — ILEMATOZOA   IN   FROGS. 

H^EMATOZOA    IN    MAN    (MALARIA). 

IN  1880  Laveran,  in  Algiers,  noticed  the  existence  of  peculiar 
structures  in  the  blood  of  a  patient  suffering  from  malaria,  and 
his  researches  were  communicated  to  the  Academy  of  Medicine  in 
Paris  in  1881  and  1882,  and  subsequently  published  in  extenso  in  a 
treatise  on  the  subject. 

Laveran  described  various  bodies  which  he  was  led  to'  regard  as 
different  stages  in  the  life-history  of  the  same  micro-parasite.  The 
most  striking  forms  were  cylindrical  elements  with  pointed  extre- 
mities. They  were  crescent -shaped  and  pigmented  in  the  middle. 
There  were  other  forms,  more  frequently  found,  which  were  either 
free  in  the  serum  or  in  contact  with  the  red  blood-corpuscles. 
They  were  more  or  less  spherical,  pigm exited,  and  endowed  with 
amoeboid  movement.  Other  forms,  again,  were  provided  with  motile 
filaments  three  or  four  times  as  long  as  the  diameter  of  a  red  blood- 
corpuscle.  And,  lastly,  there  were  little  masses  of  hyaline  material, 
which  Laveran  regarded  as  dead  forms. 

These  observations  at  first  attracted  little  attention  ;  but  they 
have  since  been  confirmed  and  extended  by  Richard,  Councilman  and 
Abbot,  Marchiafava  and  Celli,  Golgi,  Sternberg,  Osier,  the  author, 
Vandyke  Carter,  Manson,  and  others,  and  their  importance  fully 
recognised. 

The  different  forms  assumed  by  the  haematozoon  of  malaria  may 
be  described  in  two  groups  :  those  within  the  red  blood-corpuscles, 
and  those  free  in  the  serum. 

Intra-corpuscular  bodies. — These  are  of  three  kinds.     First t 

589 


590 


APPENDICES. 


structureless  protoplasmic  bodies  much  smaller  than,  and  within 
or  attached  to,  the  red  blood- corpuscles  (Fig.  233).  These  rapidly 
change  their  shape,  exhibiting  amoeboid  movement.  They  were 
first  described  by  Marchiafava  and  Celli,  and  possibly  represent  the 
first  stage  in  the  life -history  of  the  hsematozoon.  Marchiafava  and 
Celli  suggested  the  name  Plasmodium  malarise.  Second,  minute 


FIG.  233.— NON-PIGMENTED  AMEBOID  FoEMS  (Marchiafava  and  Celli). 

masses  of  finely  granular  or  of  hyaline  protoplasm  enclosing  granules 
of  pigment  (Fig.  234).  These  forms  are  sometimes  present  in  large 
numbers,  and  at  other  times  can  be  found  only  with  difficulty. 
They  are  more  or  less  spherical,  but  exhibit  amoeboid  movement, 
and  rapidly  change  their  form.  The  pigment  granules  are  also  in 
active  movement.  There  may  be  one  or  more  of  these  amoeboid 


FlG.  234.— PlGMENTED  AM(EBOID  FORMS  (Golgi). 

bodies  to  a  blood-corpuscle,  and  they  vary  in  size  ;  one  may  occupy 
the  whole  of  the  corpuscle.  In  cases  of  pernicious  malaria,  similar 
bodies  may  be  seen,  in  tissue  sections,  in  the  corpuscles  filling  the 
capillaries.  Third,  forms  which  appear  like  isolated  grains,  and 
larger  homogeneous  bodies  surrounded  by  clear  spaces  which  change 
in  outline. 

Extra-corpuscular  bodies.— These  are  the  most  striking,  and 


FIG.  235.— SEMI-LUNAR  BODIES  OF  LAVERAN  (Golgi). 

perhaps  the  most  interesting,  forms.     First,  the  semi-lunar  bodies 
Laveran.     These  are  crescent -shaped  bodies,  sometimes   pointed 


ANIMAL   MICRO-PARASITES. 


591 


at  the  extremities,  but  more  usually  rounded  off  (Fig.  235).  They 
are  not  always  curved ;  some,  indeed,  are  almost  spherical,  and 
others  sausage-shaped.  They  are  motionless.  In  many  specimens 
a  delicate  line  is  visible  on  the  concave  side  of  the  crescent  connect- 
ing the  extremities.  On  careful  examination  this  is  found  to  be 


01 


.€> 


O 


FIG.  236.—  ROSETTE  FORMS  WITH  SEGMENTATION  (Golgi). 


the  edge  of  a  very  delicate  membrane.  The  body  is  composed  of 
homogeneous  protoplasm.  Centrally  placed  is  a  collection  of  pigment 
granules,  which  on  careful  examination  can  be  distinctly  seen  to  be 
in  movement.  The  semi-lunar  bodies  vary  in  number  in  different 
cases.  Sometimes  several  can  be  seen  in  the  field  at  the  same  time, 
and  in  other  cases  they  are  only  observed  after  a  long  and  patient 
search.  They  are,  as  a  rule,  free  in  the  serum  ;  but  they  have  also 
been  seen  within  the  red  blood-cells.  Second,  finely  granular  masses 
of  protoplasm,  which  arise,  according  to  Golgi,  from  the  intra- 
oorpuscular  pigmented  bodies.  The  pigment  is  collected  in  a  rosette, 
and  the  protoplasm  by  segmentation  gives  rise  to  a  number  of  small 


FIG.  237. — FLAGELLATED  FORMS  (Vandyke  Carter). 

1.  A  flagellated  spherule ;  a,  the  same  in  the  interior  of  a  phagocyte ;  6,  free 
motile  filaments. 

spherical  forms,  which  are  ultimately  set  free  (Fig.  236).  Golgi 
believes  that  these  changes  occur  in  definite  relation  to  the  develop- 
ment of  the  paroxysm.  Third,  spherical,  pear-shaped,  or  ovoid 
bodies,  rather  smaller  than  the  red  blood-corpuscles,  and  provided 
with  one  or  more  actively  motile  flagella  (Fig.  237).  These  flagella 


592  APPENDICES. 

are  long  lash-like  filaments,  which  by  their  activity  set  the  neigh- 
bouring blood- corpuscles  in  motion.  Free  filaments  in  active  move- 
ment have  also  been  observed.  Fourth,  small  spherical  pigmeiited 
bodies  about  one-quarter  the  size  of  a  red  blood-corpuscle,  which 
exhibit  amoeboid  movement. 

Inoculation  experiments. — Marchiafava  and  Celli  assert  that 
inoculation  of  a  healthy  subject  with  blood  containing  the  parasites 
will  produce  a  paroxysm  of  ague  with  development  of  the  haematozoa. 
The  pathogenic  power  of  these  parasites,  however,  has  not  been 
established.  There  has  been  no  cultivation  of  the  parasite  outside 
the  animal  body,  and  reproduction  of  the  disease  with  a  pure  culti- 
vation. In  favour  of  its  being  a  pathogenic  organism,  Laveran 
points  out  its  invariable  presence  in  some  form  or  other  in  cases  of 
malaria ;  the  marked  changes  it  effects  in  the  red  blood-cells ;  the 
increase  in  the  number  of  the  parasites  in  proportion  to  the  severity 
of  the  attack  ;  and,  lastly,  their  disappearance  after  the  admini- 
stration of  quinine.  Others,  again,  have  doubted  the  parasitic 
nature  of  these  bodies,  and  have  looked  upon  them  as  representing 
pathological  changes  in  the  blood- cells. 

Laveran  first  of  all  suggested  the  name  Oscillaria  malarias  ;  but 
subsequently  he  recognised  that  these  bodies  belonged  to  the  animal, 
not  to  the  vegetable,  kingdom.  Osier  has  suggested  that,  tempo- 
rarily at  any  rate,  the  organism  should  be  placed  in  the  genus 
Hsematomonas  of  Mitrophanow,  thus  :  "  Genus,  Hsematomonas ; 
species,  Hsematomonas  malaria.  Definition — Body  plastic ;  ovoid 
or  globose  ;  no  differentiation  of  protoplasm,  which  contains  pigment 
grains ;  flagella  variable,  from  one  to  four ;  highly  polymorphic, 
occurring  in  (1)  amoeboid  form,  (2)  crescents,  encysted  form,  (3) 
sporocysts,  (4)  cellular  free  pigmented  bodies." 

EXAMINATION  OF  THE  HAEMATOZOA  OF  LAVERAN. 

In  the  Living  Condition. — Select  a  patient  by  preference  who  has  had 
several  attacks  of  malaria,  and  is  markedly  ansemic.  Examine  before  the 
invasion  of  the  febrile  paroxysm.  Take  two  perfectly  clean  cover-glasses 
and  two  clean  slides  ;  wash  one  of  the  fingers  of  the  patient  with  soap 
and  water,  and  then  cleanse  with  alcohol  ;  apply  a  ligature,  and  with  a 
clean  needle  puncture  the  thin  skin  near  the  root  of  the  nail  ;  touch  the 
drop  of  blood  which  collects,  with  a  clean  slide  ;  cover  quickly  with  a 
cover-glass,  and  gently  press  it  if  the  layer  of  blood  be  too  thick. 
Examine  with  a  &  o.  i. 

In  Stained  Preparations.— Puncture  the  finger  again  if  necessary  ; 
touch  the  droplet  of  blood  with  a  clean  cover-glass  ;  apply  another  cover- 
glass  ;  press  them  gently  together,  and  then  slide  them  apart  ;  stain  with 


ANIMAL   MICRO-PARASITES.  593- 

two  or  three  drops  of  alcoholic  solution  of  methylene-blue  ;  wash  off 
excess,  and  examine  in  water,  or  allow  the  preparation  to  dry,  and  mount 
in  balsam. 

H^EMATOZOA    OF    BlRDS. 

According  to  Danilewsky,  birds  suffer  from  malaria  in  both  an 
acute  and  a  chronic  form.  The  haematozoa  are  very  similar  to 
those  found  in  malaria  in  man,  and  any  slight  difference  may  be 
attributed  to  the  different  character  of  the  blood  in  birds.  Grassi 
and  Feletti  have  described  two  kinds  of  malarial  hsematozoa  in 
birds,  one  kind  belonging  to  the  genus  Hcemamveba  and  the  other 
to  the  genus  Laverania. 

H^MATOZOA    OF   TURTLES. 

Danilewsky  has  also  minutely  described  and  figured  hsematozoa  in 
the  blood  of  turtles,  which  in  some  stages  of  their  life  history  very 
closely  resemble  those  found  by  Laveran  in  man. 

H^MATOZOA   OF    EQUINES   AND    CAMELS    (SURRA    DISEASE.) 

Swra  is  a  blood  disease  occurring  in  horses,  mules,  and  camels,, 
characterised  by  fever  accompanied  by  jaundice,  petechise  of  mucous 
membranes,  great  prostration,  and  rapid  wasting  terminating  in 
death.  The  average  duration  of  the  disease  is  about  two  months. 
No  organic  lesions  are  found  after  death,  but  a  parasite  exists  in 
the  blood  during  life.  By  means  of  subcutaneous  inoculation,  and 
by  the  introduction  into  the  stomach  of  blood  containing  the 
parasite,  the  disease,  according  to  Evans,  can  be  transmitted  to 
healthy  animals.  The  importance  of  this  disease  may  be  realised 
from  the  fact  that  on  one  occasion  in  India  the  3rd  Punjab  Cavalry 
lost  no  less  than  three  hundred  horses  from  it. 

The  disease  has  not  been  observed  to  be  contagious  or  infectious 
in  the  ordinary  sense,  but  the  possibility  of  its  conveyance  by  means 
of  large  brown  flies  has  been  suggested.  These  flies  attack  the 
horses  so  vehemently  that  the  blood  frequently  streams  from  the 
bites  ;  and  the  opinion  that  they  propagate  the  disease  is  prevalent 
among  the  natives.  At  the  same  time  it  has  been  particularly 
noted  that  where  the  disease  has  broken  out  the  water  was  very 
impure. 

Evans  discovered  a  haematozoon,  in  1880,  in  all  the  diseased  horses 
and  mules  examined  ;  in  all  diseased  camels,  with  one  exception ;  and 
in  the  dogs  which  had  been  subjected  to  experimental  inoculations 

38 


594  APPENDICES. 

Evans  stated  that  when  he  first  discovered  the  parasite  he 
thought  it  was  a  spirillum,  but  very  speedily  on  closer  examination 
arrived  at  an  opposite  opinion. 

To  him  the  organism  presented  the  appearance,  when  fresh  and 
active,  of  an  apparently  round  body,  tapering  in  front  to  form  a 
neck  and  terminating  in  a  blunt  head.  Posteriorly  he  described  a 
tapering  tail,  from  which  there  extended  a  long  slender  lash.  At 
the  head  end  there  appeared  in  one  or  two  cases  a  circlet  of 
pseudopods,  and  as  the  body  slowly  died  in  serum  it  gave  the 
appearance  of  flattening  out.  After  watching  very  closely  all  its 
•changes  of  form  and  movements,  Evans  came  to  the  conclusion  that 
there  existed  on  either  side  of  the  body  two  fin-like  papillae,  one 
near  where  the  neck  began  an(J  the  other  close  to  where  the  tail 
began.  In  only  very  few  instances  he  was  able  to  see  the  four  at 
once.  He  suggested  that  these  processes  were  of  the  nature  of 
pseudopods. 

The  parasite  he  described  as  extremely  active  in  its  movements, 
with  an  undulatory,  eel-like  motion,  progressing  for  the  most  part 
head-end  foremost,  but  occasionally  moving  in  the  direction  of  the 
lash  when  tugging  at  a  corpuscle.  In  fresh  blood  these  organisms 
resembled  spermatozoa  in  colour ;  but  their  peculiar  characteristic 
was  the  power  they  possessed  of  attacking  and  disintegrating  the 
red  corpuscles. 

Occasionally  two  were  observed  to  unite  and  swim  off  as  one 
body ;  but  the  mode  of  union  was  a  disputed  point.  Evans  thought 
that  they  joined  with  their  respective  heads  and  tails  in  the  same 
direction,  overlapping  each  other;  but  others  to  whom  they  were 
shown  were  of  opinion  that  they  fastened  with  their  tails  in 
opposite  directions. 

The  parasites  were  not  always  present  in  the  blood,  but  were 
•observed  to  come  and  go  in  successive  broods.  Evans  referred  the 
organism  to  Lewis  for  his  opinion  as  to  its  nature.  Lewis  arrived 
at  the  conclusion  that  the  parasite  was  "  more  nearly  related  to 
that  which  he  found  in  the  blood  of  rats  than  to  any  other  " ;  but 
he  was  of  opinion  at  the  time  that  they  did  not  appear  exactly 
the  same. 

Five  years  later  Surra  broke  out  in  British  Burma,  and  Steel 
was  deputed  to  investigate  the  outbreak.  Steel  confirmed  the 
communicability  of  the  disease  to  dogs,  horses  and  mules  by 
ingestion  and  inoculation,  but  he  considerably  supplemented  Evans' 
views  as  to  the  nature  of  the  disease  by  careful  thermometric 
observations  :  these  finally  led  him  to  regard  the  disease  as  a  true 


ANIMAL   MICRO-PARASITES.  595 

relapsing  fever,  closely  resembling  relapsing  fever  in  man.  At  the 
same  time  it  is  worth  recording  that  until  Steel  observed  the 
presence  of  the  parasite  described  by  Evans  he  regarded  the  out- 
break as  malarious  in  origin,  and  provisionally  termed  it  gastric 
typhoid.  In  the  Burma  outbreak,  as  in  the  Punjab  epidemic, 
considerable  evidence  was  adduced  in  favour  of  regarding  the  disease 
as  being  due  to  bad  water  supply. 

Steel  succeeded  in  staining  the  organism  with  aniline  dyes,  but 
his  description  of  the  parasite  in  the  frssh  state  differs  very 
materially  from  that  given  by  Evans. 

Steel  failed  to  recognise  the  round  body  tapering  in  front  to  a 
neck.  To  him  the  bodies  appeared  thick  in  the  middle,  gradually 
diminishing  in  size  in  either  direction,  with  a  blunt  and  rigid 
extremity  at  one  end.  The  opposite  end  he  described  as  tapering 
in  such  a  way  as  to  produce  a  subspiral  prolongation,  which  was 
uncurled  and  lashed  about  freely  like  a  whip.  This  tail  was 
described  as  slender  in  relation  to  the  general  size  of  the  parasite ; 
but  under  the  highest  power  available  the  presence  of  a  colourless 
nagellum  could  not  be  detected,  nor,  he  adds,  did  the  movements  of 
the  blood -constituents  indicate  its  existence. 

Steel  also  failed  to  see  the  slightest  sign  of  the  two  fin -like  papillae 
on  each  side  as  described  by  Evans — an  opinion  in  which  he  was 
supported  by  Lewis. 

These  two  observers,  Evans  and  Steel,  also  differed  as  to  whether 
the  movement  could  be  called  spiral.  Steel  felt  convinced  that  their 
movement  was  as  much  of  that  nature  at  times  as  can  be  expected 
from  organisms  with  so  open  a  corkscrew  shape ;  while  Evans 
maintained  an  opposite  view.  In  the  dried  and  stained  specimens 
Steel  observed  that  they  retained  their  subspiral  form  of  body  and 
markedly  spiral  form  of  tail. 

Steel  found  that  the  disease  could  be  communicated  to  the  dog 
and  to  the  monkey,  and  then  discussed  the  resemblance  of  the  parasite 
to  the  spirillum  of  relapsing  fever  in  man. 

From  the  different  appearances  presented  by  the  parasite  when 
in  the  living  state  and  when  dried  and  stained,  Steel  thought  that 
there  was  probably  a  still  closer  resemblance  to  the  living  spirillum 
than  to  the  dried  and  stained  one,  and  argued  that  the  figures  of 
spirilla  like  corkscrews  must  be  purely  imaginary.  Steel,  it 
must  be  observed,  founded  these  remarks  upon  figures  in  text- 
books, and  not  on  photographs  or  on  a  practical  acquaintance 
with  the  spirillum  of  relapsing  fever.  One  cannot  refrain  from 
pointing  out  the  value  of  photomicrographs,  for  they  cannot  be 


596  APPENDICES. 

called  into  question;  and  had  Steel  studied  photographs  of 
spirilla  he  would  not  have  regarded  the  corkscrew  appearance  as 
imaghiM  ry. 

Steel  found  the  parasite  in  all  cases,  and  further  observed  that 
it  appeared  as  the  temperature  rose  and  disappeared  during  the 
apyrexial  periods. 

From  all  these  observations  Steel  concluded  as  follows : — That 
relapsing  fever  of  mules  is  an  invariably  fatal  disorder,  characterised 
by  the  periodical  occurrence  of  attacks  of  high  fever,  during  which  a 
special  organism  closely  resembling  the  spirillum  of  relapsing  fever 
in  man  is  found  in  the  blood.  This  organism  is  one-sixth  the  size  of 
a  red  corpuscle  in  width  and  three  to  six  times  in  length.  It  is 
eel-like,  and,  when  dried  and  stained,  presents  a  thick  portion — the 
body — and  a  spiral  tail.  The  latter  takes  less  of  the  dye  than  the 
former,  and  commences  as  a  sudden  narrowing  of  the  body,  termi- 
nating by  a  fine  point.  This,  he  insisted,  had  nothing  of  the  nature 
of  an  infusorian  nagellum.  The  thick  portion  tapers  in  either 
direction  from  its  centre,  and  terminates  in  front  abruptly  in  a  rigid 
process,  with  probably  some  holdfast  organ.  The  sharpness  of  the 
head  end  varies  in  different  animals.  The  body  portion  he  described 
as  spiral,  and  so  closely  in  general  appearances  to  resemble  the 
spirillum  of  relapsing  fever  that  he  concluded  that  the  organism 
was  undoubtedly  a  spiral  bacterium  and  named  it  after  its  discoverer 
Spirochceta  Evansi.  This  view,  however,  would  not  be  accepted  by 
Evans,  who  maintained  that,  whatever  it  might  be,  it  was  not  a 
member  of  the  family  of  bacteria. 

In  the  face  of  these  conflicting  opinions  Evans,  in  1885,  submitted 
to  the  author  preparations  of  the  organism  in  the  blood  as  well 
as  material  from  the  lungs  and  intestines  of  a  camel  that  had 
succumbed  to  the  disease. 

On  examining  a  stained  preparation  the  author  found  that 
with  a  power  of  200  diameters  a  number  of  the  parasites  could 
be  distinguished  in  the  field  of  the  microscope,  and  with  TV  and 
Y\r  o.  i.  objectives  the  individual  characteristics  were  clearly  brought 
out.  These  were  quite  sufficient  at  once  to  dispel  the  idea  of  its  being 
a  spirillum.  It  was  obvious  that  it  was  a  more  highly  organized 
micro-parasite,  presenting  very  peculiar  and  distinctive  structural 
appearances. 

The  author  came  to  the  following  conclusions  : — 

The  somewhat  tapering  central  portion,  or  body,  of  the  parasite 
is  continuous  at  one  end  with  a  whip-like  lash,  and  at  the  other  end 
terminates  in  an  acutely-pointed  stiff  filament  or  spine-like  process. 


ANIMAL   MICRO-PARASITES.  597 

Here  and  there,  possibly  from  injury  or  want  of  development,  the 
spine-like  process  appears  to  be  blunted  or  absent.  By  very  careful 
focussing  on  the  upper  edge  of  the  central  portion,  the  author 
discovered  the  existence — much  more  markedly  in  some  of  the 
parasites  than  in  others — of  a  longitudinal  membrane  with  either 
a  straight  or  undulating  margin.  The  membrane  is  attached  along 
the  body,  arising  from  the  base  of  the  rigid  filament,  and  becomes 
directly  continuous  at  the  opposite  end  with  the  flagellum.  In  some 
cases  the  edge  only  is  deeply  stained,  giving  the  appearance  of  a 
thread  continuous  with  the  flagellum,  so  that  one  might  be  easily 
led  to  overlook  the  membrane,  and  imagine  that  the  flagellum  arose 
from  the  opposite  end  of  the  body,  at  the  base  of  the  spine-like 
process. 

Close  to  the  base  of  the  spine-like  process  a  clear  unstained  spot 
is  in  many  parasites  easily  distinguished ;  and  at  the  opposite  end 
there  is,  in  some,  the  appearance  of  the  deeply-stained  protoplasmic 
contents  having  contracted  within  the  faintly-stained  membranous 
investment.  When  the  longitudinal  membrane  has  a  wavy  outline 
the  undulations  are  much  more  marked  in  some  cases  than  in  others. 
Here  and  there  the  wavy  outline  appears  first  on  the  one  side  of 
the  central  portion  and  then  on  the  other ;  but  there  never  is  any 
waving  outline  on  both  sides  of  the  same  part  of  the  body,  and  this 
was  explained  by  a  careful  examination,  which  showed  that  the 
somewhat  ribbon -like  parasite  had  become  doubled  on  itself.  The 
discovery  of  this  undulating  membrane  at  once  suggested  to  the 
author  an  explanation  of  the  lateral  pseudopodia  described  by 
Evans.  If  we  imagine  that  we  are  looking  down  upon  the  parasite, 
with  the  edge  of  the  membrane  towards  us,  one  can  conceive  that 
the  rapid  undulations,  first  on  one  side  and  then  on  another, 
might  give  an  image  upon  the  retina  which  could  be  construed 
as  due  to  the  protrusion  of  lateral  pseudopodia.  In  stained 
preparations  no  trace  of  the  jcirclet  of  pseudopods  could  be 
discovered,  and  the  undulating  membrane  may  account  for  this 
appearance  also. 

Owing  to  the  somewhat  curved  and  twisted  shape  of  the  parasite 
and  the  curling  of  the  flagellum  in  the  stained  preparations,  it 
was  difficult  to  make  exact  measurements ;  but  the  average  width, 
according  to  whether  the  membrane  was  visible  or  not,  varied  from 
1  to  2  /x,  and  the  length  of  the  body  from  20  to  30  /M.  The  flagellum 
was  about  the  same  length  as  the  body. 

Here  and  there  in  a  stained  preparation  there  were  the  forms 
already  described  by  Evans  resulting  from  the  fusion  of  two  para- 


598 


APPENDICES. 


sites.  But  the  union  obviously  took  place  by  the  noil- flagellated 
ends,  for  the  two  flagella  were  frequently  turned  in  the  same 
direction,  so  that  the  fused  parasites  resembled,  as  Evans  sub- 
sequently suggested,  a  trophy  of  buffalo  horns.  Here  and  there 
more  than  two  parasites  had  united,  forming  a  stellate  group ;  and 
in  one  case  the  author  noticed  that  the  individuals  had  apparently 
united  with  their  non-flagellated  ends  just  overlapping,  so  that  the 
unstained  spot  in  one  was  just  situated  in  a  line  with  the  unstained 
spot  of  the  other. 

In  Evans's  Report,  Lewis's  opinion  is  given  that  these  parasites 
differed  slightly,  but  still  were  closely  allied  to  certain  flagellated 
organisms  which  had  been  observed  by  him  in  rats  in  India.  On 


FIG.  238.— "SURRA"  PARASITES  OCCURRING  SINGLY  AND  FUSED. 
(From  preparations  stained  with  magenta,    x   1200.    Lent  by  Dr.  Evans.) 

referring  to  his  original  memoir,  it  will  be  found  that  his  description 
and  woodcut  differed  very  materially  from  the  Surra  parasite  as 
just  described,  though  a  microphotograph  which  Lewis  had  appended 
to  the  memoir  after  it  was  written,  indicated  a  great  similarity 
to  this  organism.  To  the  author  the  organisms  appeared  not  only 
closely  allied,  but,  as  far  as  one  can  judge  from  figures  and  descrip- 
tions, morphologically  identical  with  the  parasites  described  by 
Mitrophanow  in  the  carp,  and  as  a  matter  of  fact,  instead  of  a 
mere  resemblance,  the  rat  and  the  Surra  parasites,  when  stained, 
are  found  to  be  morphologically  identical. 


•-, 


ANIMAL   MICRO-PARASITES.  599 

H^MATOZOA    OF    RATS. 

In  describing  these  organisms,  Lewis  remarked  that  it  wa& 
strange  that  they  had  not  occupied  attention  before,  and  suggested 
as  an  explanation  that  possibly  European  rats  did  not  harbour  these 
parasites.  The  author  examined  a  few  white  rats,  but  without 
success,  and  then  proceeded  to  examine  the  blood  of  common  brown 
rats,  trapped  from  the  London  sewers,  and  discovered  that  these 
organisms  are  to  be  found  in  no  less  than  25  per  cent,  of  apparently 
healthy  animals.  The  first  question  which  naturally  arose  was 
whether  these  organisms  in  European  rats  were  identical  with  those 
described  by  Lewis  in  Indian  rats. 

If  we  refer  to  the  description  given  by  Lewis,  we  find  that  he 
states  that  when  he  first  noticed  them  he  thought  they  were  vibrios 
or   spirilla,     The  drop  of   blood 
under   examination  appeared   to 
quiver  with  life ;  and  on  diluting 
the  blood,  motile  filaments  could 
be    seen     rushing     through    the 
serum    and    tossing    the    blood- 
corpuscles  about  in  all  directions. 

The  filaments  were  pale  and 
translucent,  without  any  trace 
of  visible  structure  or  granu- 
larity, and  they  were  more  un- 
dulatory  in  movement  than  FlG.  239.._pARASITE8  IN  THK  BLOOD 
spirilla.  A  corpuscle  might  be  OP  RATS  (Lewis), 

observed    to     quiver,     and    this 

could  be  distinctly  traced  to  be  due  to  the  existence  of  a  flagellum, 
apparently  a  posterior  flagellum,  as  the  organisms  seemed  generally 
to  move  with  the  thicker  end  forward;  no  flagellum  could  be 
detected  at  the  opposite  end.  The  greater  number  of  the  figures- 
in  the  woodcut  (Fig.  239)  are  described  as  representing  these 
organisms  a  few  hours  after  the  blood  had  been  obtained,  when 
their  movements  are  not  so  rapid,  and  the  flagellum  becomes  more 
easily  recognisable. 

This  observation  led  Kent,  who  named  the  organism  Herpetomonas 
Lewisi,  to  remark  that  if,  as  Lewis  is  inclined  to  maintain,  that 
organ  "  propels  instead  of  draws  the  animalcule  through  the  in- 
habited serum,  we  have  presented  a  structural  and  functional  feature 
without  parallel  among  the  other  representatives  of  these  Protozoa 
flagellata,  the  recognition  of  which  would  demand  the  creation  of  a 


APPENDICES. 

distinct  generic  and  family  group  for  the  reception  of  these  singular 
organisms."  In  his  later  paper,  however,  Lewis  came  to  the  con- 
elusion  that,  like  the  generality  of  flagellated  organisms,  the  rat 
parasites  moved  with  the  lash  in  front. 

On  careful  examination  the  plasma  which  constituted  the  thicker 
portion  of  their  substance  was  observed  to  suddenly  swell  out  so  as 
to  divide  the  body  into  two  parts,  as  seen  in  the  centre  of  the  figure ; 
.at  other  times  two  or  three  such  constrictions  or  dilatations  were 
detected,  and  at  other  times  the  body  assumed  an  arrow  shape,  as 
depicted  at  the  lower  part  of  the  figure.  When  dried,  and  stained 
with  a  little  weak  solution  of  aniline-blue,  the  body  presented  a  very 
different  appearance.  It  was  found  to  have  contracted  irregularly, 
and  to  manifest  a  somewhat  granular  and  shreddy  appearance, 
.suggestive  of  a  coagulated  fibro-albuminous  substance.  The  body 
portion  became  flattened  towards  its  middle  to  double  its  original 
width,  and  both  ends  almost  acutely  pointed,  while  the  flagellum 
was  only  partly  visible.  After  fixing  with  osmic  acid  they  measured 
•0*8  to  1  ju,  in  width, 'and  20  to  30  ft  in  length  ;  the  flagellum  was  about 
as  long  as  the  body :  so  that  the  total  length  of  the  organism  was 
.about  50  fj..  Lewis  detected  these  parasites  in  29  per  cent,  of  the 
.species  Mus  decumanus  and  Mus  rufescens,  but  failed  to  find  them 
in  mice.  He  considered  that  they  had  many  features  in  common 
with  motile  organisms  of  vegetable  origin ;  but  they  appeared  to 
approach  much  more  closely  to  the  Protozoa,  more  particularly 
several  of  the  species  of  Dujardin's  Cercomonas.  He  points  out 
that  many,  however,  believe  that  these  organisms  are  zoospores  and 
not  animalcules.  To  him  they  also  seemed  to  be  not  unlike  the 
flagellated  parasite  described  by  Biitschli. 

The  latter  observer  detected  flagellated  organisms  (Leptomonas 
liutschlii)  in  the  intestinal  canal  of  a  free  nematode  (Trilobus 
gracilis).  They,  too,  form  stellate  colonies,  like  the  Surra  parasite, 
owing  to  their  being  attached  by  their  non- flagellated  ends.  When 
detached  from  these  colonies  they  presented  a  somewhat  spindle- 
shaped  body  about  11  ft  in  length,  with  a  somewhat  thick  flagellum 
about  double  this  length,  so  that  the  total  length  of  the  protozoon 
would  be  33  /w,,  or,  as  Lewis  states,  about  half  the  length  of  the 
flagellated  organism  in  the  rat's  blood.  Near  the  base  of  the 
flagellum,  Butschli's  protozoon  presented  a  contractile  vacuole, 
but  Lewis  was  unable  to  detect  any  such  vacuole  in  the  rat 
haematozoa. 

In  conclusion,  Lewis  observed  that  very  probably  these  organ- 
isms corresponded  with  the  vermicides  observed  by  Goss  in  the 


ANIMAL   MICRO-PARASITES.  601 

blood  of  a  field  mouse,  and  he  also  mentions  that  Chaussat  found 
minute  "  nematodes  "  in  the  blood  of  a  black  rat. 

Wittich  discovered  in  the  blood  of  hamsters  whip- like  bodies  with 
lively  movements.  They  resembled  frog's  spermatozoa,  possessing 
a  thick  portion  continued  into  a  long  lash -like  thread.  Wittich 
considered  them  identical  with  the  organisms  described  by  Lewis,  and 
they  also  were  observed  in  apparently  healthy  animals.  Koch  later 
met  with  the  same  organisms. 

Like  Lewis,  the  author  found  that  the  blood  of  the  common 
brown  rat  in  England  appeared  to  quiver  with  life,  and  that  the 
parasites  were  extremely  difficult  to  examine  until  their  movement 
was  arrested  for  a  moment  or  they  became  imprisoned  in  the  serum 
areas.  After  examining  with  various  powers,  from  a  -I  dry 
to  a  ^v  o.  i.  of  Powell  and  Lealand,  the  author  came  to  the 
following  conclusion : — That  they  are  polymorphic,  presenting  for 


FIG.  240.— A  MONAD  IN  RAT'S  BLOOD.  The  organism  is  represented  at  partial 
rest  with  its  posterior  filament  impinging  on  a  corpuscle,  and  showing  the 
undulating  longitudinal  membrane,  the  long  flagellum,  find  the  refractive 
spherules  in  the  granular  protoplasm  (  x  3000). 

the  most  part  slightly  tapering  bodies  which  terminate  at  one  end  in 
a  stiff,  immotile,  acutely-pointed  flexible  filament  or  spine-like  process, 
and  at  the  opposite  end  are  provided  with  a  long  flagellum,  while, 
longitudinally  attached,  a  delicate  undulating  fin-like  membrane  can 
be  traced,  which  starts  from  the  base  of  the  posterior  filament,  and 
becomes  directly  continuous  with  the  flagellum  (Fig.  240). 

With  careful  illumination  the  body  is  found  to  be  distinctly 
granular,  with  one  or  more  highly -refractive  spherules.  When  the 
rapid  movement  is  arrested  the  undulating  membrane  is  distinctly 
visible.  The  best  opportunity  occurs  for  seeing  this  when  the 
organism  comes  to  partial  rest  with  its  stiff  filament  against  a 
corpuscle,  as  if  to  obtain  a  point  cFappui,  while  lashing  its  flagellum 
in  all  directions  (Fig.  241,  b).  At  other  times,  when  the  parasite  has 
impinged  with  its  posterior  extremity  against  a  corpuscle,  or  the 
stiff  filament  is  apparently  entangled  in  debris,  the  movements  of 
the  organism  give  one  the  idea  of  its  endeavouring  to  set  itself  free. 


602 


APPENDICES. 


but  the  author   has  not  been  able  to  persuade  himself  that  they 
"  attack  and  disintegrate  "  the  red  blood- corpuscles. 

In  the  active  state  the  thicker  portion,  or  body,  appears  to 
twist  and  bend  from  side  to  side  with  great  activity.  The  organism 
can  turn  completely  round  with  lightning  rapidity,  so  that  the 
flagellum,  at  one  moment  lashing  in  one  direction,  is  suddenly 
observed  working  in  the  opposite  direction.  Then  suddenly  the 
organism  makes  progression,  and  it  can  be  distinctly  seen  to  move 
in  the  direction  of  the  flagellum,  the  flagellum  threading  its  way 
between  the  corpuscles  and  drawing  the  rest  of  the  organism  after 
it.  Currents  set  up  by  evaporation  may  undoubtedly  here  and 
there  produce  the  appearance  of  the  organism  "wriggling  along" 
with  its  flagellum  posterior ;  but  the  author  was  convinced,  after 


FIG.  241.— MONADS  IN  RAT'S  BLOOD,  x  1200.  a,  A  monad  threading  its  way  among 
the  blood-corpuscles  ;  b,  another  with  pendulum  movement  attached  to  a  cor- 
puscle ;  c,  angular  forms  ;  d,  encysted  forms  ;  e  and  /,  the  same  seen  edgeways. 

hours  of  patient  observation,  that  in  the  normal  mode  of  progression 
the  flagellum  acts  as  a  tractellum  and  not  as  a  pulsellum.  By 
treating  cover-glass  preparations  with  osmic  acid  the  appearances 
obtained  are  very  similar  to  those  shown  in  Lewis's  photographs, 
so  that  there  is  no  doubt,  in  spite  of  the  descriptions  not  completely 
according,  that  they  are  one  and  the  same  organism.  There  was  a 
great  likeness  to  the  organisms  described  by  Mitrophanow,  and  to  the 
Surra  parasite ;  and  when  the  author  had  stained  the  rat  parasites, 
the  closest  examination  confirmed  his  belief  that  they  were  morpho- 
logically identical  with  the  stained  parasites  of  Surra. 

Cover-glasses  with  a  thin  layer  of  blood  may  be  passed  three 
times  through  the  flame  of  a  Bunsen  burner  in  the  way  commonly 
employed  for  examining  micro-organisms,  and  stained  with  an 


ANIMAL    MICRO-PARASITES.  603 

aqueous  solution  of  fuchsin,  methyl-violet,  or  Bismarck- brown, 
or  with  aurantia,  nigrosin,  and  other  aniline  dyes.  The  following 
method  will,  however,  be  found  most  instructive: — Use  freshly 
prepared  saturated  solution  of  fuchsin  or  methyl-violet  in  absolute 
alcohol,  and  put  a  drop  with  a  pipette  on  the  centre  of  the  prepara- 
tion ;  do  not  disturb  the  drop-form  for  a  few  moments  ;  then,  before 
the  alcohol  has  evaporated,  wash  off  the  excess  of  stain.  It  will  be 
found  that  where  the  drop  rested  the  organisms  will  be  very  deeply 
stained,  while  in  the  surrounding  area  the  colour  will  vary  in 
intensity.  By  the  effect  of  the  different  degrees  of  staining  much 


FIG.  242.— MONADS  IN  KAT'S  BLOOD  STAINED  WITH  METHYL  VIOLET,  SHOWING 
MEMBRANE  UNDER  DIFFERENT  ASPECTS,  BLOOD-CORPUSCLES,  SOME  CRE- 
NATED  AND  STAINED  DlSCS  (  X  1200). 

may  be  learnt  (Fig.  242).  In  one  organism  the  body  and  entire 
membrane  will  be  equally  stained ;  in  another  the  margin  of  the 
membrane  only.  In  some  the  posterior  stiff  filament  is  stained, 
and  at  its  base  a  darkly  stained  speck  is  very  striking ;  and  in 
other  cases,  again,  the  posterior  filament  is  only  faintly  tinged,  or 
an  unstained  spot  occurs  near  its  base. 

H^EMATOZOA    OF   FlSH. 

In  the  year  1883  Mitrophanow  published  a  paper  in  which  he 
gave  an  account  of  organisms  in  the  blood  of  the  mud-fish  and  the 
carp. 

In  the  blood  of  the  mud-fish  (Cobitis  fossilis)  the  organisms  at 
the  first  glance  looked  like  minute  nematodes,  but  the  appearances, 
and  changes  which  took  place  on  further  examination  showed 
nothing  in  common  with  worms  (Fig.  243).  As  a  1  per  cent,  salt 
solution  had  been  added  to  the  blood  under  examination,  it  occurred 


604  APPENDICES. 

to  Mitrophanow  that  they  were  possibly  the  cytozoa  described  by 
Gaule ;  but  this  idea  was  dismissed  by  the  fact  that  they  were 
found  in  blood  to  which  110  salt  solution  was  added.  Their  size 
varied  from  30  to  40  /*  in  length  and  1  to  1|  //,  in  width.  At  first 
their  rapid  movements  baffled  examination,  but  as  the  rapidity 
lessened  there  was  the  appearance  of  a  curling  movement  in  the 
body  portion  and  a  swinging  movement  of  the  lash.  The  organism 
moved  in  the  direction  of  the  lash,  the  anterior  end  of  the  body 
being  more  pointed  than  the  posterior,  and  gradually  fining  off  into 
the  lash.  When  the  body  seemed  to  rest,  the  lash  might  be  seen  to 


FIG.  243.— ORGANISMS  IN  THE  BLOOD  OF  MUD-FISH  (Hfemcitomoncts  colitis),  a,  First 
Variety ;  6,  second  variety ;  c,  third  variety,  d,  First  variety  in  a  state  of 
diminished  activity,  e,  The  same  after  treatment  with  osmic  acid.  (Mitro- 
phanow. ) 

whip  out  in  all  directions.  As  the  movement  of  the  body  gradually 
diminished,  it  appeared  to  have  a  complicated  screw  form,  the  axis 
of  the  screw  corresponding  to  the  body  to  which  an  undulating 
membrane  is  fastened  spirally.  This  could  be  distinguished  when 
the  organism  was  dying,  because  the  body  in  death  contracted,  and 
the  membrane  then  looked  like  a  spiral  addition.  Thus  the 
organism  consisted  of  a  body,  a  spiral  membrane,  and  a  flagellum. 

With  higher  magnification  the  organism  appeared  to  consist  of  a 
refractive,  strongly  contractile  protoplasmic  substance,  which,  when 
death  occurred,  formed  a  shapeless  mass.  In  the  same  blood  two 
other  forms  were  observed :  one  without  a  membrane,  but  having 


ANIMAL   MICRO-PARASITES.  G05 

two  highly  refractive  spherules  in  the  protoplasm ;  and  another  with 
neither  membrane  nor  flagellum,  consisting  of  very  granular  proto- 
plasm with  several  refractive  spherules,  and  capable  of  protruding 
processes  like  pseudopodia. 

In  the  carp  (Fig.  244)  the  parasite  is  perceptibly  larger,  and 
possesses  an  undulating  membrane  fastened  along  the  edge  of  the 
long  body.  When  the  body  bent  first  towards  one  side  and  then  to 
the  other,  a  wave-like  movement  was  observable  at  the  free  edge  of 
this  membrane. 

These  parasites  were  found  in  all  the  mud-fish  examined  except 


FIG.  244. — ORGANISMS  IN  THE  BLOOD  OF  THE  CARP. 

a,  I,  c,  Ha'iiMtomonas  carassii  ;  d,  e,  /,  g,  h,  other  organisms  in  the  same  blood 
(Mitrophanow). 

one,  and  in  greater  numbers  in  the  hot  months.  In  the  carp 
tlx-y  were  only  found  occasionally.  Mitrophanow  described  other 
varieties,  which  he  considered  were  possibly  not  complete  organisms, 
but  developmental  forms.  He  considered  that  these  organisms  were 
infusoria  between  the  genera  Cercomonas  and  Trichomonas,  with 
.luvat  similarity  to  the  Trichomonas  described  in  the  LieberkulmV 
glands  of  fowls  and  ducks  (Eberth). 

On  account  of  their  special  habitat,  Mitrophanow  suggested  a 
new  genus — Haematomonas,  defining  this  genus  as  follow*. :-— Parasites 
of  normal  fish-blood,  worm-like,  actively  moving  organisms,  with 
indistinct  differentiation  of  body  parenchyma.  l>odii*>  pointed  at 


f,06  APPENDICES. 

both  ends,  30  to  40  p,  long  and  1  to  1|  /*  wide.     May  possess  in 
front  a  nagellum,  and  on  one  side  an  undulating  membrane. 
Species  : — 

Hsematomonas  cobitis. — Body  provided  with  a  spiral  membrane 
and  a  nagellum  at  the  fore-end.  Parenchyma  of  body  homogeneous. 
Second  variety,  body  and  nagellum  only.  Movement  undulatory, 
body  containing  highly  refractive  spherules.  Third  variety,  plasma- 
like  body,  without  membrane  or  nagellum ;  quickly  changes  form 
by  sending  out  processes  laterally,  and  contains  two  to  four  refractive 
spherules.  Blood  of  mud-fish. 

Hsematomonas  carassii.— Long  bodies,  with  narrow  membrane 
attached  along  the  whole  length;  less  actively  motile.  Several 
forms  also  observed  strikingly  smaller  than  the  above ;  many  disc- 
shaped.  Often  seen  attached  to  a  red  corpuscle,  setting  them  in 
motion  by  their  movements.  Blood  of  carp. 

The  morphological  identity  of  the  rat  and  Surra  parasites  has  been 
established  by  the  author,  and  both  saem  morphologically  identical 
with  the  organism  of  Mitrophanow.     If  we  follow  Mitrophanow,  we 
must  obviously  enlarge  his  genus  of  Hsematomonas.    The  author  doss 
not  agree  with  Mitrophanow  in  the  advisability  of  adopting  this 
entirely  new  generic  name.     Mitrophanow  suggested  this  new  term 
because  of  the  special  habitat,  normal  fish-blood,  of  the  species  he 
discovered.    But  the  characteristic  features  of  these  organisms  are  the 
characteristic  marks  of  the  genus  Trichomonas.     It  seems,  therefore, 
that  they  are  embraced  by  the  old  genus  Trichomonas,  and  that  there 
is  no  need  to  create  a  new  one — Hsematomonas.    The  common  habitat 
of  these  species  may  be  expressed  by  grouping  them  together  in  one 
sub-genus — Trichomonas  sanguinis  ;  but  the  question  arises  whether 
they  are  distinct  species.     If  it  were  not  for  the  different  description 
given  by  Mitrophanow  of  the  organism  in  the  mud-fish,  the  author 
would  be  inclined  to  say  that  all  these  organisms  belonged  to  one  and 
the  same  species,  which  might  well  be  named  Trichomonas  sanguinis. 
The  monad  in  the  rat  and  the  Surra  parasite  are  morphologically 
identical  with  each  other,  and  both,  as  far  as  one  can  judge  from 
the  description,  morphologically   identical   with   the  monad  in  the 
blood  of  the  carp.     We  have,  however,  seen  that  the  organism  in 
Surra  is  believed  to  be  pathogenic,  and  too  much  stress  must  not  be 
laid  on  morphological  identity.     There  is  strong  evidence  in  favour 
of  believing  in  its  pathogenic  properties;  but  at  the  same  time  it 
must  be  borne  in  mind  that  the  organism  has  never  been  isolated 
apart  from  the  blood,  and  the  disease  then  produced  by  its  introduc- 
tion into  healthy  animals.     It  is  quite  possible  that  the  parasites  in 


ANIMAL   MICRO-PARASITES.  607 

Surra  are  only  associated  with  the  disease,  the  impoverished  blood 
affording  a  suitable  nidus  for  their  development,  while  the  con- 
taminated water  may  be  the  common  source  of  the  organism  and  of 
the  disease.  On  the  other  hand,  the  organism  in  the  rat  is  found  in 
apparently  perfectly  healthy,  well-nourished  animals.  The  author 
suggests  that  the  parasites  observed  in  the  rat  and  hamster  should 
be  named  after  Lewis,  Trichomonas  Lewisi',  the  organism  in  the 
mule,  camel  and  horse  after  its  discoverer,  Trichomonas  Evansi', 
and  that  the  names  Trichomonas  cobitis  and  Trichomonas  carassii 
should  be  substituted  for  the  names  of  the  species  described  by 
Mitrophanow.  Thus  we  should  have  added  provisionally  to  the 

Genus— TRICHOMONAS. 

Sub-genus — Trichomonas  sanguinis.  Definition  :  Elongated 
tapering  bodies,  provided  with  a  spiral  (T.  cobitis),  or 
longitudinal  (T.  carassii,  Lewisi,  Evansi)  membrane,  ter- 
minating in  a  rigid  filament  and  an  anterior  flagellum. 
Highly  polymorphic.  Habitat,  the  blood. 

Species. — Trichomonas      cobitis     (Hcem,atomonas      cobitis 

Mitrophanow) — Mud-fish. 
Trichomonas    carassii    (tfcematomonas    carassii 

Mitrophanow) — Carp. 
Trichomonas     Lewisi     (Herpetomonas      Lewisi 

Kent) — Rat,  hamster. 
Trichomonas        Evansi — (Spirochceta       Evansi 

Steel) — Horse,  mule,  camel ;  (pathogenic  ?). 


H^MATOZOA    OF   THE    FROG. 

Lankester  described  an  organism  which  he  had  discovered  in  the 
blood  of  the  frog  (Rana  esculenta).  It  consisted  of  a  minute  pyri- 
form  sac,  with  the  narrower  end  bent  round  on  itself  somewhat 
spirally,  and  the  broader  end  spread  out  into  a  thin  membrane, 
which  exhibited  four  or  five  folds  and  was  prolonged  on  one  side  into 
a  very  long  flagellum.  The  wall  of  the  sac  was  striated,  nucleated 
and  granular ;  the  membrane  undulated  during  life,  and  the 
flagellum  was  also  motile.  It  was  named  Undulina  ranarum,  but 
subsequently  recognised  as  idnetical  with  Tnjpanosoma  sanguinis 
described  by  Gruby.  In  the  same  blood  Lankester  also  discovered 
little  oblong  bodies,  in  many  cases  attached  to  the  end  of  the  red 
corpuscles,  and  suggested  a  geneticai  connection  with  the  Undulina. 


608  APPENDICES. 

One  or  more  motionless  filaments  were  occasionally  observed  attached 
to  these  bodies.  Gaule  subsequently  observed  the  same  bodies,  and 
regarded  them  as  resulting  from  the  metamorphosis  of  the  cells  of 
the  frog's  blood.  Gaule's  observations  were  refuted  by  Lankester  in 
1882,  the  parasitic  nature  insisted  upon,  and  the  organism  named 
Drepanidium  ranarum.  Lankester  suggested  that  they  were 
probably  the  young  stage  of  a  sporozoon  allied  to  Sarcocystis  or  to 
Coccidium 


APPENDIX    III. 

PSOROSPERMS   OR   COCCIDI A.—  AMOEBA  COLI. 

PSOROSPERMS    OR    COCCIDIA. 

GREYISH- WHITE  nodules  may  occasionally  be  found  in  the  liver  of  a 
rabbit,  the  result  of  a  disease  which  may  be  mistaken  for  tuber- 
culosis. This  disease  often  proves  fatal,  and  may  occur  in  an 
epidemic  form  in  rabbit  warrens.  The  nodules  have  cheesy  or 
purulent  contents,  which  are  found,  on  microscopical  examination, 
to  contain  great  quantities  of  Coccidium  oviforme. 

The  coccidia  pass  from  the  intestine  into  the  bile-ducts.  The 
walls  of  the  bile -ducts  become  dilated  and  folded  ;  and  irregular 
cavities  result  from  the  partial  or  complete  disappearance  of  the 
dividing  walls  of  the  altered  ducts.  The  folds  are  composed  of 
connective  tissues  lined  with  columnar  epithelium,  and  the  coccidia, 
in  different  stages  of  development,  are  found  between  the  cells,  and 
free  in  the  cavities  of  the  nodules. 

The  individual  coccidia  are  egg-shaped  bodies.  They  possess  a 
thick  smooth  shell,  with  an  opening,  or  micropyle,  at  one  end,  and 
protoplasmic  contents  which  may  completely  fill  the  capsule  or  be 
collected  into  a  spherical  mass. 

After  passing  from  the  liver  and  intestine,  -these  oval  bodies 
undergo  a  further  development.  According  to  Leuckart,  who  has 
very  fully  described  this  parasite,  the  protoplasmic  contents  divide 
into  four  masses,  and  from  each  is  developed  a  C-shaped  hyaline 
rod,  the  cavity  of  which  is  occupied  by  closely  packed  granules.  In 
this  condition  they  remain  until  they  gain  access  to  a  fresh  host. 

Coccidium  oviforme  has  been  found  in  the  human  liver,  and  also 
in  sheep,  dogs,  and  cats.  Similar,  but  not  identical,  bodies  occur  in 
mice,  and  also  in  fish  and  other  cold-blooded  animals. 

Miescher's  tubes  are  peculiar  structures  found  in  swine,  cattle, 
sheep,  deer,  and  mice.  They  consist  of  a  firm  envelope  inclosing  a 
number  of  reniform  or  bean-shaped  bodies. 

609  39 


610  APPENDICES. 

Pfeiffer's  bodies.— Pfeiffer  has  described  certain  appearances 
which  he  attributes  to  coccidia,  in  epithelial  cells  in  small-pox, 
vaccinia,  and  other  vesicular  diseases.  They  are  probably  only 
derived  from  the  cell  nucleus,  and  are  not  parasites. 

Canc&r  bodies. — In  sections  of  malignant  growths  stained  by 
aniline  dyes,  certain  bodies  have  been  found  and  minutely  described 
and  figured  by  various  investigators,  and  a  causal  relation  sug- 
gested. Darier  first  described  bodies  like  cysts,  with  spores,  in 
Paget's  disease  of  the  nipple.  Wickham  found  similar  structures 
and  figured  them.  Nils  Sjobring  described  a  cancer  parasite,  and 
illustrated  his  researches  with  plates.  Russell  drew  attention  to 
certain  bodies  in  cancerous  tumours,  with  a  great  affinity  for 
fuchsine.  Soudakewitch,  Podwyssozki,  Sawtschenko,  Ruffer,  and 
Walker  have,  among  others,  contributed  to  the  literature  of  the  so- 
called  cancer  parasites.  These  bodies  appear  in  the  form  of  ref ractile 
spherical  elements,  which  stain  well  with  reagents,  such  as  the 
Ehrlich-Biondi  stain.  Sections  are  left  in  this  stain  for  twenty-four 
hours,  washed  in  alcohol,  cleared  in  xylol,  and  mounted  in  xylol 
balsam.  The  spherical  bodies  have  sometimes  a  radiate  appearance. 
These  bodies  have  not  been  cultivated,  and  inoculation  experiments 
with  cancerous  tissue  have  been  negative.  The  opinion  is  now  very 
generally  held  that  these  bodies  are  not  parasites,  but  that  changes 
occur  in  the  cells  and  nuclei,  resulting  in  the  formation  of  peculiar 
structures,  which  have  been  brought  to  light  by  the  use  of  aniline 
dyes  and  complex  staining  methods.  We  are  justified  in  concluding 
that  the  cause  of  cancer  is  unknown. 

Ballance  and  Shattock  have  made  repeated  attempts  to  cultivate 
parasitic  protozoa  from  malignant  tumours,  and  they  have  extended 
their  researches  to  vaccinia  and  molluscum  contagiosum,  but  with 
negative  results.  Sand  and  water  were  used  as  the  medium  for 
these  experiments.  Cultivations  were  made  from  nine  scirrhous 
carcinomata  of  the  breast,  five  sarcomata  from  different  sources,  two 
melanotic  sarcomata  from  horses,  and  a  sarcoma  from  a  dog.  In 
every  instance  the  result  was  negative.  No  traces  of  protozoic  life 
could  be  found,  in  spite  of  examinations  at  regular  intervals,  and 
repeated  for  periods  of  many  months. 

AMCEBA  COLI. 

Lbsch,  Grassi,  Kartulis,  and  others  have  described  an  amoeba  in 
the  intestines  of  patients  suffering  from  dysentery.  Losch  adminis- 
tered the  fresh  dejecta  of  a  patient  containing  the  amoeba?  to  dogs, 


ANIMAL   MICRO-PARASITES.  611 

and  in  one  case  a  mucous  mass  was  passed  containing  a  number 
of  amoebae.  Eighteen  days  afterwards  the  dog  was  killed,  and  the 
mucous  membrane  of  the  intestine  was  reddened,  swollen,  and 


m 

"(0 


FIG.  245.— AMCEBA  COLI  IN  INTESTINAL  Mucus  (Loscn). 

ulcerated  in  three  places.  The  mucus  in  the  rectum  and  in  the 
ulcers  contained  numerous  amoebae.  Cunningham,  who  has  found 
the  amoebae  in  choleraic  and  other  cases,  and  in  the  intestine  of  the 
cow  and  horse,  does  not  attach  any  importance  to  their  presence. 


APPENDIX    IV. 

APPARATUS,   MATERIAL,  AND   REAGENTS   EMPLOYED 
IN  A  BACTERIOLOGICAL  LABORATORY. 

(A)    HlSTOLOGICAL   APPARATUS. 

Microscope. — For  the  investigation  of  micro-organisms  a  good 
microscope  with  oil-immersion  system  and  a  condenser,  such  as 
Abbe's,  is  essential.  Such  instruments  are  supplied  by  Zeiss,  Leitz, 
Reichert  &  Hartnack  in  Germany,  and  Powell  &  Lealand,  Swift 
&  Baker  in  England.  Zeiss  supplies  a  micrometer  eyepiece,  with 
-directions  for  use.  Some  such  arrangement  is  essential  for  the 
measurement  of  bacteria.  Other  accessories  to  the  microscope  are  : 

A  large  bell-glass,  for  covering  the  microscope  when  not  in  use. 

About  a  foot  square  of  blackened  plate-glass. 

A  white  porcelain  slab  of  the  same  size. 

Glass  bottles,  with  ground -glass  stoppers,  for  alcoholic  solutions  of 
aniline  dyes,  etc. 

Glass  bottles,  with  funnels,  for  aqueous  solutions  of  the  dyes,  and 
-others  provided  with  pipettes. 

A  small  rod-stoppered  bottle  of  cedar  oil.  This  is  recommended  by 
Zeiss  in  preference  to  other  oils  for  his  immersion  lenses. 

Set  of  small  glass  dishes  or  capsules  and  watch-glasses,  for  section- 
staining,  etc. 

Stock  of  best  glass  slides,  in  packets  of  fifty. 

Several  boxes  of  round  and  square  thin  cover-glasses,  in  various  sizes, 
of  the  best  quality. 

Needle-holders,  with  a  couple  of  platinum  needles,  and  a  packet  of 
ordinary  sewing-needles. 

Glass  rods  drawn  out  to  a  fine  point ;  useful  for  manipulating  sections 
when  acids  are  employed. 

Platinum  or  plated  copper  section-lifters. 

One  pair  of  small  brass  or  spring-steel  platinum-pointed  forceps,  for 
holding  cover-glasses. 

One  pair  of  brass  tongg. 

612 


APPARATUS,   MATERIAL,    AND  REAGENTS.  61$ 

Collapsible  tubes,  for  containing  Canada  balsam  ;  very  serviceable  for 
transport  and  general  use. 

Turn-table  for  sealing  cover-glass  preparations,  with  rings  of  cement. 

Boxes  for  preparations,  book-form. 

Tickets  and  labels,  various  sizes. 

Soft  rags  or  old  pocket-handkerchiefs,  for  removing  cedar  oil  from 
immersion  lens,  cleaning  cover-glasses,  etc. 

Chamois  leather  for  wiping  lenses. 

Warm  Stages. — In  addition  to  those  already  described,  Schafer 
and  Strieker  have  constructed  warm  stages  for  accurate  observations. 
Schafer's  apparatus  consists  of  a  vessel  (/),  filled  with  water  which 
has  been  boiled  to  expel  the  air,  and  heated  by  means  of  a  gas-flame 
at  g.  The  warmed  water  ascends  the  indiarubber  tube  (c)  to  the 


FIG.  246.— SCHAFER'S  WARM  STAGE. 

brass  box  (a).  The  box  is  pierced  by  a  tubular  aperture  to  admit 
light  to  the  object,  and  has  an  exit  tube  (c'),  by  which  the  cooled 
water  from  the  stage  returns  to  be  reheated  by  the  flame  g.  At  d 
is  a  gas-regulator,  so  that  a  constant  temperature  at  any  desired 
point  can  be  maintained. 

Strieker's  stage,  in  which  warm  water  or  steam  can  be  used  for 
heating,  and  by  the  employment  of  iced  water  also  used  for  observing 
the  effects  of  cold,  is  shown  in  Fig.  247.  It  consists  of  a  hollow 
rectangular  box,  with  a  central  opening  (C)  permitting  the  passage 
of  light.  The  water  makes  its  exit  and  entrance  at  the  side  tubes 
(a,  a),  and  the  temperature  is  indicated  by  a  thermometer  in  front. 


614 


APPENDICES. 


A  more  complicated  apparatus,  combining  both  a  warm  stage 
and  a  gas  chamber,  is  shown  in  Fig.  248.  This  consists  of  a  rect- 
angular piece  of  ebonite  (E  E)  fixed  to  a  brass  plate  which  rests 
on  the  stage  of  the  microscope.  On  the  upper  surface  of  the  ebonite 
is'another  brass  plate  (P),  with  an  aperture  (G)  leading  into  a  brass 


FIG.  247.—  STEICKEK'S  WARM  STAGE. 

tube  closed  below  by  a  piece  of  glass.  To  heat  the  apparatus  the 
copper  wire  B  is  placed  on  the  tube  a,  and  its  extremity  heated  by 
the  flame  of  the  lamp.  The  nearer  the  lamp  to  the  stage  the  higher 
the  temperature,  which  is  indicated  by  the  thermometer  (t).  To 


J> 


FIG.  248.— STRICKEB'S  COMBINED  GAS  CHAMBER  AND  WARM  STAGE. 

employ  it  as  a  gas  chamber  the  wire  B  is  laid  aside,  and  the  gas 
is  conducted  into  the  chamber  by  the  tube  a',  and  escapes  by  the 
tube  a. 

Microtome — Schanze's  is   much   in   favour  in  Germany,   but 
Jung's  of  Heidelberg,  though  a  somewhat  cumbrous  instrument,  is 


APPARATUS,   MATERIAL,   AND   REAGENTS.  615 

preferred  by  many  workers.     Smaller  accessories,  which  should  be 
within  reach,  are — 

A  small  can  of  sewing-machine  oil. 

A  soft  rag  and  chamois  leather,  for  wiping  the  knives  immediately 
after  use. 

Stone  and  leather,  for  setting  and  sharpening  the  same. 
Two  or  three  camel's-hair  brushes. 

A  freezing  microtome  is  very  useful :  such  as  Swift's,  which  is 
used  by  the  author;  and  the  method  of  embedding  in  celloidin  is 
combined  with  the  ordinary  process  of  freezing. 


(B)  REAGENTS  AND  MATERIAL  EMPLOYED  IN  THE  PROCESSES  or 
HARDENING,  DECALCIFYING,  EMBEDDING,  FIXING  AND  CUTTING 
OF  TISSUES. 

Alcohol,  absolute. 
Bergamot  oil. 

Celloidin. 

Dissolved  in  equal  parts  of  ether  and  alcohol. 

Cork,  or  stock  of  ready-cut  corks. 

Ebner's  solution.     A  mixture  in  the  following  proportions  : — 

Hydrochloric  acid  ....  '5 

Alcohol ,100 

Distilled  water 20 

Chloride  of  sodium          ...  5 

Formalin. 

Gelatine. 

Melted  in  a  small  porcelain  capsule,  and  set  aside  ready  to   be 
re-melted  when  required  for  use. 

Glycerine  gelatine  (Klebs). 

Best  well- washed  gelatine      .         .         .  .10 

Add  distilled  water,  allow  gelatine  to  swell  up,  pour 

off  excess  of  water,  melt  gelatine  with  gentle  heat, 

add 
Glycerine       .....  .10 

Lastly,  a  few  drops  of  phenol  for  preservation. 

Gum. 


616  APPENDICES. 

Kleinenberg's  solution. 

Saturated  watery  solution  of  picric  acid          .         .100 
Strong  sulphuric  acid    ...          .          .          .          2 

Filter,  and  add 

Distilled  water 300 

Miiller's  fluid. 

Bichromate  of  potash  .  .          .  ...  2 

Sulphate  of  sodium  .  .          .  .          .          .  1 

Distilled  water      .  .  ...  .         ...  100 

Osmic  acid. 

Distilled  water       .         .         •....«        -•         •          .100 
Osmic  acid     .         .          .         .          .          .  .      '.          .  -5 

Paper  trays  (or  small  glass  capsules). 

Paraffine. 

Spermaceti. 

Xylol. 

(C)  REAGENTS  FOR  EXAMINING  AND  STAINING  MICROSCOPICAL 
PREPARATIONS. 

1.  Acetic  acid,  strong. 

2.  Alcohol,  absolute. 

3.  Alcohol,  60  per  cent. 

4.  Alcohol,  acidulated. 

Alcohol     .....  100 

Hydrochloric  acid       ...  1 

5.  Alum  Carmine  (Grenadier). 

Carmine    .         .         .  j 

Five  per  cent,  solution  of  alum  .         .         .100 

Boil  twenty  minutes,;  filter  when  cold. 

6.  Ammonia,  strong. 

7.  Aniline. 

8.  Aniline  water. 

Distilled  water          .  100 

Aniline    .  K 

•  •       .       •  .  •  ••    v1 ...         y 

Shake  well,  and  filter  emulsion. 


APPARATUS,   MATERIAL,   AND   REAGENTS.  617 

9.  Bismarck-brown. 

(a)  Concentrated  solution  in  equal  parts  of  glycerine  and  water. 

(b)  Aqueous  solution. 

Bismarck-brown  .  .  .  .  .  2 
Alcohol  .  .  .  .".  .  .  .  .15 
Distilled  water 85 

10.  Borax-carmine  (Grenacher). 

Borax 2 

Carmine  ........          5 

Distilled  water         .         .         .         .         .         .100 

To  the  dark  purple  solution  add  a  5  per  cent,  solution  of  acetic 
acid  until  a  red  colour  is  produced  ;  set  aside  twenty-four  hours ; 
filter,  and  add  a  drop  of  carbolic  acid. 

11.  Cedar  oil. 

12.  Ehrlich-Biondi  solution  (Heidenhain). 

To 

Saturated  aqueous  solution  of  Orange.  G.  .100 

Add 

Saturated  aqueous  solution  of  Rubin.  S.  .  .  20 
„  „  „  Methyl-green.  OO.  50 

To  the  mixture         ....  1 

Add  water 100 

13.  Eosin. 

(a)  Saturated  alcoholic  solution. 

(b)  Aqueous  solution. 

Distilled  water.         ....  .100 

Eosin       .         .         ....  5 

14.  Ether. 

15.  Fuchsine. 

(a)  Saturated  alcoholic  solution. 

(b)  Aqueous  solution. 

Fuchsine .... 

Alcohol .15 

Water      .....  .85 

16.  Gentian- violet. 

(a)  Saturated  alcoholic  solution. 

(6)  Aqueous  solution. 

Gentian-violet.         .  .  2 -25 

Distilled  water  .     100 


OP  THH 

1  UNIVERSITY 


618  APPENDICES. 

17.  Gibbes'  solution,  for  double  staining. 
Take  of 

Rosaniline  hydrochlorate .....          2 

Methylene-blue        ......          1 

Triturate  in  a  glass  mortar. 

Dissolve  aniline  oil  .  .         .          .         .         3 

In  rectified  spirit      ...         .         .          .         .15 

and  add  slowly  to  the  above. 

Lastly,  slowly  add  distilled  water      .         .          .15 
Keep  in  stoppered  bottle. 

18.  Glycerine,  pure. 

19.  Hsematoxylin  solution. 

Hsematoxylin  .         .         .          .          .         .          .         2 

Alcohol 100 

Distilled  water          .         .         .         .         .         .100 

Glycerine          .......     100 

Alum       ........         2 

20.  Iodine  solution. 

Iodine,  pure      .......  1 

Iodide  of  potassium  ......  2 

Distilled  water 50 

21.  Iodine  solution  (Gram). 

Iodine      .....  1 

Iodide  of  potassium .         .         .                  .         .         2 
Distilled  water 300 

22.  Lithium-carmine  solution  (Orth). 

Saturated  solution  of  carbonate  of  lithium         .     100 
Carmine  ......  .         2'5 

23.  Magenta  solution  (Gibbes). 

Magenta ......  2 

Aniline  oil        ...                  .  3 

Alcohol  (sp.  gr.  -830) 20 

Distilled  water          ....  20 

24.  Methylene-blue. 

(a)  Concentrated  alcoholic  solution. 

{b)  Aqueous  solution. 

Methylene-blue         ....  2 

Alcohol 15 

Water     .         .  85 


APPARATUS,    MATERIAL,    AND   REAGENTS.  619 

•(c)  Koch's  solution. 

Concentrated  alcoholic  solution  of  methylene-blue         1 
Ten  per  cent,  potash  solution    .... 
Distilled  water 200 

(d)  Loffler's  solution. 

Concentrated  alcoholic  solution  of  methylene-blue       30 
Solution  of  potash,  1  to  10,000  .         .         .     100 

25.  Methyl-violet. 

(a)  Concentrated  alcoholic  solution. 

(b)  Aqueous  solution. 

Methyl- violet  .         .         .         .         .         .         .         2 -2 5 

Distilled  water          ...         .         .         .100 

(c)  Koch's  solution. 

Aniline  water  .          .          .         .         .         .         .100 

Alcoholic  solution  of  methyl-violet    .  .11 

Absolute  alcohol       .         .         .         .         .         .10 

26.  Neelsen's  solution. 

Dissolve  f uchsine      ..... 

In  alcohol 10 

Add  a  5  per  cent,  watery  solution  of  carbolic  acid     100 

27.  Nitric  acid,  pure. 

28.  Orseille  (Wedl). 

Dissolve  pure  ammonia-free  orseille  in 

Absolute  alcohol       ...  .20 

Acetic  acid       ....  5 

Distilled  water          .  •       40 

until  a  dark  red  liquid  results.     Filter. 

29.  Picric  acid. 

(a)  Concentrated  alcoholic  solution. 

(b)  Saturated  aqueous  solution. 

30.  Picro-carmine  (Ranvier). 

Carmine  . 

Distilled  water          .  .10 

Solution  of  ammonia  3 

Triturate ;  add  cold  saturated  solution  of  picric 
acid  ....     200 


620  APPENDICES. 

31.  Picro-lithium-carmine  (Orth). 

To  above-mentioned   lithium-carmine  solution 

add  saturated  solution  of  picric  acid        .          .     2'3 

32.  Potash  solution. 

(a)  1  to  3     per  cent. 

(b)  10    „      „ 

(c)  33    „      „ 

33.  Safranine. 

(a)  Concentrated  alcoholic  solution. 

(b)  Watery  solution       .         .         .         .         ;     1  per  cent. 

34.  Sulphuric  acid,  pure. 

35.  Salt  solution        .     :  .'..    ,'    .    ,'   .         .     0-8  per  cent. 

36.  Turpentine. 

37.  Vesuvin. 

(a)  Concentrated  alcoholic  solution. 

(b)  Watery  solution. 

Water,  distilled. 

Water,  sterilised. 

Distilled  water  can  be  kept  for  use  in  a  wash  bottle,  or  far 
better  in  a  siphon  apparatus.  Sterilised  water  is  convenient  in 
plugged  sterile  test-tubes,  which  may  be  kept  close  at  hand  in  a 
beaker,  or  tumbler,  with  a  pad  of  cotton  wool  at  the  bottom.  The 
numbered  reagents  can  be  conveniently  arranged  on  shelves  within 
easy  reach.  Alcoholic  solutions  of  the  aniline  dyes  and  other  special 
preparations  should  be  kept  in  bottles  with  ground- glass  stoppers. 
Aqueous  solutions  of  the  dyes  may  be  kept  in  bottles  with  funnel 
niters,  and  the  solution  filtered  before  use.  To  both  aqueous  and 
alcoholic  solutions  a  few  drops  of  phenol,  or  a  crystal  of  thymol, 
should  be  added  as  a  preservative.  For  the  rapid  staining  of  cover- 
glass  preparations,  it  is  convenient  also  to  have  the  most  frequently 
used  stains  (fuchsine,  methyl- violet)  in  bottles  provided  with  pipette 
stoppers. 

(D)  REAGENTS  FOR  MOUNTING  AND  PRESERVING  PREPARATIONS. 

Acetate  of  potash. 
Concentrated  solution. 

Asphalte  lac. 


APPARATUS,   MATERIAL,   AND   REAGENTS.  621 

Canada  balsam. 
Dissolved  in  xylol. 

Glycerine  gum  (Farrant's  solution). 
Glycerine. 
Water. 

Saturated  solution  of  arsenious  acid. 
Equal  parts  ;  mix,  and  add  of  picked  gum  arabic  half  a  part. 

Hollis'  glue. 
Zinc-white. 

(E)  DRAWING  AND  PHOTOGRAPHIC  APPARATUS. 

Camera  Lucida. — The  camera  lucida  of  Zeiss  is  an  excellent 
instrument,  though  many  prefer  the  pattern  made  by  Nachet  of 
Paris.  Combined  with  the  use  of  a  micromillimeter  objective,  it 
affords  also  a  simple  method  for  the  measurement  of  bacteria. 

For  drawing  microscopical  appearances,  and  for  illustrating 
microscopical  specimens  with  or  without  the  use  of  a  camera  lucida, 
the  following  materials  should  be  within  reach  : — 

Pencils. 

Etching  pens. 

Prepared  Indian  ink. 

Water-colour  paints  and  brushes. 

Ordinary  and  tinted  drawing  paper  and  other  usual  accessories. 

Photo-micrographic  Apparatus, — Zeiss  of  Jena,  Seibert  & 
Kraft  of  Wetzlar,  Nachet  of  Paris,  and  Swift  &  Son  of  London,  may 
all  be  recommended  for  constructing  an  arrangement  in  which  the 
photographic  camera  is  combined  with  the  microscope. 

The  best  models  have  been  described  fully  in  the  chapter  on 
Photography  of  Bacteria.  The  accompanying  figure  (Fig.  249) 
illustrates  a  model  in  which  the  microscope  is  used  in  the  vertical 
position. 

For  illumination  either  sunlight  or  artificial  light  may  be  em- 
ployed. In  the  case  of  sunlight  a  heliostat  is  necessary  to  procure 
the  best  results  ;  but  as  sunlight  is  not  always  available  by  day,  and 
it  is  also  more  convenient  for  many  to  work  at  night,  it  is  better  to 
have  recourse  altogether  to  artificial  light.  Excellent  results  may 
be  obtained  with  an  ordinary  paraffine  lamp,  or  with  magnesium, 
oxycalcium,  or  electric  light. 


622 


APPENDICES. 


FIG.  249.— VEKTICAL  MICRO-PHOTOGKAPHIC  APPARATUS. 


(F)  STERILISATION  APPARATUS. 

Steam-steriliser. — A  cylindrical  vessel  of  tin  about  half  a  metre 
or  more  in  height,  jacketed  with  thick  felt,  and  provided  with  a 
conical  cap  or  lid  (Fig.  250).  The  lid  is  also  covered  with  felt,  has 
handles  on  either  side,  and  is  perforated  at  the  apex,  to  receive  a 
thermometer.  Inside  the  vessel  is  an  iron  grating  or  diaphragm 
about  two-thirds  the  way  down,  which  divides  the  interior  into 
two  chambers — the  upper  or  "  steam-chamber,"  and  the  lower  or 
"water-chamber."  A  gauge  outside  marks  the  level  of  the  water 
in  the  lower  chamber ;  this  should  be  kept  about  two-thirds  full. 


APPARATUS,   MATERIAL,   AND   REAGENTS. 


The  apparatus  stands  upon  three  legs,  and  is  heated  from  below  with 
two  or  three  Bunsen  burners,  or  a  Fletcher's 
burner.  It  is  employed  for  sterilising 
nutrient  media  in  tubes  or  flasks,  for  cooking 
potatoes,  or  hastening  the  filtration  of  agar- 
agar.  When  the  thermometer  indicates 
100°  C.  the  lid  is  removed,  and  test-tubes 
are  lowered  in  a  wire  basket  by  means  of 
a  hook  and  string,  and  the  lid  quickly  re- 
placed. Potatoes  or  small  flasks  are  lowered 
into  the  cylinder  in  a  tin  receiver  with  a 
perforated  bottom,  which  rests  upon  the- 
grating  and  admits  of  its  contents  being 
exposed  to  the  steam.  A  larger  model  is 
shown  in  Fig.  33. 

Hot-air  Steriliser. — A  cubical  chest  of 
sheet  iron  with  double  walls,  supported  on 
four  legs  ;  it  may  also  be  suspended  on  the 
wall  of  the  laboratory,  with  a  sheet  of 

FiG.250.-KocH's  STEAM-   Asbestos  intervening  (Figs.  251  and  252). 

It  is  heated  with  a  rose  gas-burner  from 
below,  and  the  temperature  of  the  interior  in- 


STERILISEE. 


clicated  by  a  thermometer  inserted  through 
a  hole  in  the  roof ;  in  a  second  opening  a 
gas  regulator  can  be  fixed.  Test-tubes, 
flasks,  funnels,  cotton  wool,  etc.,  may  be 
sterilised  by  exposure  to  a  temperature 
of  150°  C.  for  an  hour  or  more. 


FIG.  251.— HOT-AIR  STERILISER. 


FIG.  252.— SECTION  OF  HOT- 
AIR  STERILISER. 


624 


APPENDICES. 


(G)  APPARATUS  AND  MATERIAL  FOR  PREPARING  AND  STORING 
NUTRIENT  GELATINE  AND  NUTRIENT  AGAR-AGAR. 

Water-bath. —  A  water-bath  on  tripod   stand   is   required   for 

boiling  the  ingredients  of  nutrient  jellies  and  for  general  purposes. 

The  lid  may  be  conveniently   composed   of  a  series   of   concentric 

rings,  so  that  the  mouth  of  the  vessel  may  be  graduated  to  any 

size  required. 

Test-tube  Water- bath. — This  consists  of  a  circular  rack  for 

test-tubes  within  a  water-bath.     It  is  sometimes  employed  instead 

of  the  steam   cylinder  for  sterilising   nutrient   jelly   in  tubes,   by 

boiling  for  an  hour  for  three  successive  days. 

Hot- water  Filter. — A  copper 
funnel  with  double  walls,  the  inter- 
space between  which  is  filled  "with 
hot  water.  A  glass  funnel  fits  in- 
side the  copper  cone,  the  stem  of 
the  glass  funnel  passing  through 
and  being  tightly  gripped  by  a  per- 
forated caoutchouc  plug,  which  fits 
in  the  opening  at  the  apex  of  the 
cone.  The  water  in  the  cone  is 
heated  by  applying  the  flame  of  a 
burner  to  a  tabular  prolongation 
of  the  water- chamber.  In  a  more 
recent  model,  as  represented  in 
Fig.  31,  this  prolongation  is  dis- 
pensed with,  and  the  temperature  is 
maintained  by  means  of  a  circular 
burner  which  acts  at  the  same  time 
as  a  funnel  ring.  In  Rohrbeck's 
model  the  funnel  of  the  filter  is 
connected  with  a  flask,  from  which 

the  test-tubes  can  be  easily  filled  with  the  liquid  jelly  (Fig.  253). 
Glass  Vessels.— A  number  of  glass  vessels  should  be  kept  in 

stock  according  to  requirements. 

Bohemian  hard  glass  flasks  are  employed  in  several  sizes,   for 

boiling  nutrient  media.     The  conical  forms  are  especially  used  in  the 

larger  sizes  for  storing  nutrient  jelly. 

Glass  funnels,   large  and  small,  are  necessary,  not  only  in  the 

processes  of  preparing  nutrient  jelly,  but  for  filtering  solutions  of 

aniline  dyes  and  for  general  purposes. 


FIG.  253.— HOT- WATER  FILTERING 
APPARATUS  WITH  RING  BURNE  R. 


APPARATUS,   MATERIAL,   AND   REAGENTS.  625 

A  liberal  supply  of  test-tubes  should  always  be  kept  in  stock,  as 
they  are  not  only  employed  for  the  tube- cultivations,  but  can  be 
conveniently  used  for  storing  bouillon,  sterilised  water,  etc. 

Cylindrical  glasses  graduated  in  cubic  centimetres,  10  ccm.,  100 
ccm.,  500  ccm.,  are  required  for  measuring  the  liquid  ingredients 
of  nutrient  jelly,  and  also  in  preparing  the  various  staining 
.solutions. 

A  large  wide- mouthed  glass  jar,  with  a  glass  cover,  is  extremely 
useful.  It  must  be  padded  at  .the  bottom  with  cotton  wool  for 
containing  a  stock  of  tubes  of  sterilised  nutrient  jelly,  and  should 
be  placed  within  reach  on  the  working  table. 

Balance  and  Weights.— A  balance,  with  large  pans  and  set  of 
gramme  weights,  is  constantly  required. 

Cotton  Wool. — The  best  or  "  medicated  "  cotton  wool  should  be 
procured.  , 

Gelatine. — The  gelatine  for  bacteriological  purposes  must  be  of 
the  very  best  quality  (gold  label). 

Agar-agar. — This  is  also  called  Japanese  Isinglass ;  it  consists 
of  the  shrivelled  filaments  of  certain  Algse  (Gracilaria  lichenoides 
and  Gigartina  speciosa). 

Peptonum  Siccum. 

Table  Salt. — Prepared  table  salt  can  be  obtained  in  tins  or 
packets. 

Litmus  Papers. — Blue  or  red  litmus  paper  in  cheque-books,  for 
testing  the  gelatine  mixture,  etc. 

Carbonate  of  Soda. — A  bottle,  containing  a  saturated  solution 
of  carbonate  of  soda,  and  provided  with  a  pipette  stopper,  may  be 
kept,  especially  for  use  in  the  preparation  of  nutrient  jelly. 

Lactic  Acid. 

Filter  Paper. — For  filtering  gelatine,  stout  Swedish  filter  paper 
of  the  best  quality  is  recommended. 

Flannel  or  Frieze. — This  is  employed  as  a  substitute  for,  or 
combined  with,  filter  paper  in  the  preparation  of  nutrient  agar- 
agar. 


(H)  APPARATUS  FOR  EMPLOYMENT  OF  NUTRIENT  JELLY  IN  TEST-TUBE 
AND  PLATE-CULTIVATIONS. 

Wire  Cages. — These  cages  or  crates  are  used  for  containing 
test-tubes,  especially  when  they  are  to  be  sterilised  in  the  hot-air 
steriliser  ;  or  for  lowering  tubes  of  nutrient  jelly  into  the  steam- 
steriliser,  etc.  (Fig.  254). 

40 


626 


APPENDICES. 


Test-tube  Stands.— The  ordinary  wooden  pattern,  or  the 
metallic  folding  stands,  are  called  into  use 
for  holding  cultivations.  Pegged  racks  are 
also  recommended  for  draining  test-tubes 
after  washing. 

Caoutchouc  Caps. — These  are  caps  for 
fitting  over  the  cotton -wool  plugs,  and  may 
be  used  in  different  sizes  for  test-tubes  and 
stock-flasks. 

Platinum  Needles. — A  platinum 
needle  for  inoculating  nutrient  media,  ex- 
amining cultivations,  etc.,  consists  of  two  or 
three  inches  of  platinum  wire  fixed  to  the 
end  of  a  glass  rod.  Several  of  these  needles  should  be  made  with 
platinum  wire  of  various  thicknesses.  A  piece  of  glass  rod,  about 
seven  inches  long,  is  heated  at  the  extreme  point  in  the  flame  of 


FIG.  254.— WIRE  CAGE 
FOB  TEST-TUBES. 


FIG.  255. — PLATINUM  NEEDLES;  STRAIGHT,  HOOKED,  LOOPED. 

a  Bunsen  burner,  and  a  piece  of  platinum  wire,  held  near  one 
extremity  with  forceps,  is  then  fused  into  the  end  of  the  rod. 
Some  needles  should  be  perfectly  straight,  and  kept  especially  for 


FIG.  256.— DAMP  CHAMBER  FOR  PLATE-CULTIVATIONS. 

inoculating  test-tubes  of  nutrient  jelly.  For  other  purposes  the 
needles  may  be  bent  at  the  extremity  into  a  small  hook,  and 
)thers  provided  with  a  loop  (Fig.  255). 


APPARATUS,   MATERIAL,   AND   REAGENTS. 


627 


Tripod  Levelling-stand. — A  triangular  wooden  frame  sup- 
ported upon  three  screw-feet  which  enable  it  to  be  raised  or  lowered 
to  adjust  the  level. 


FIG.  257. — APPARATUS  EMPLOYED  FOR  PLATE-CULTIVATIONS. 

Tripod  Stand ;  Glass  Dish,  filled  with  cold  or  iced  water ;  Sheet  of  Plate-glass  ; 
Spirit  Level,  and  Glass  Bell. 

Large   Glass   Plate. — A   piece  of  plate-glass,   or   a   pane   of 
ordinary  window-glass,  about  a  foot  square. 

Spirit  Level. 

Glass  Bells  and  Dishes. — Shallow  glass  bells  and  dishes,  for 
making  a   dozen  or  more    damp    chambers 
(Fig.  256),  and  for  completing  the  apparatus 
for  pouring  out  liquefied   nutrient  jelly  on 
glass  plates  or  slides  (Fig.  257). 

Iron      Box A     box     of     sheet-iron 

(Fig.  258),  for  containing  glass  plates  during 
their  sterilisation  in  the  hot-air  steriliser, 
and  for  storing  them  until  required  for  use. 

Glass  Plates. — Small  panes  of  glass, 
about  six  inches  by  four.  Not  less  than 
three  dozen  are  required  for  a  dozen  damp 
chambers. 

Glass  Benches. — These  are  necessary  for  arranging  the  glass- 
plates  or  slides  in  tiers  in  the  damp  chambers  (Fig.  256).      Metal 


FIG.  258.— Box  FOR 
GLASS  PLATES. 


FIG.  259.— GLASS  BENCHES  FOR  GLASS  PLATES  OR  SLIDES. 

shelves   may  be    substituted    for  them,    but   the  former  are  to  be 
preferred.     They  can  be  easily  made,  in  any  number  required,  by 


APPENDICES. 


cementing  a  little  piece  of  plate-glass  at  either  end  of  a  glass 
.slip  (Fig.  259). 

Glass  Rods. — One  dozen  or  more  glass  rods,  twelve  to  eighteen 
inches  in  length.  They  are  employed  for  smoothly  spreading  out 
the  liquefied  nutrient  gelatine  or  agar-agar  on  the  glass  plates,  etc. 

Thermometers. — Two  or  three  centigrade  thermometers. 


(I)  APPARATUS  FOR  PREPARATION  OF  POTATO-CULTIVATIONS. 

Israel's  Case. — Sterilising  instruments  in  the  flame  of  a  Bunsen 

.burner  is  most  destructive.  It  is  better,  therefore,  to  have  a  sheet- 
iron  case  (Fig.  260)  to 
contain  potato-knives, 
scalpels  and  other  in- 
struments, and  to  ster- 
ilise them  by  placing 
the  case  in  the  hot-air 
steriliser  for  an  hour 
at  150°  C.  The  box 
can  be  opened  at  the 
FIG.  260.— ISRAEL'S  CASE.  side,  and  each  instru- 

ment   withdrawn    with 

a  pair  of  sterilised  forceps  when  required  for  ase 

Glass  Dishes.— Several  shallow  glass  dishes  are  required  for 

preparing  damp  chambers  for  potato-cultivations  (Fig.  261).     The 

upper,  being  the  larger,  fits 

over   the  lower,  and  having 

no   handle,    admits  of   these 

damp  chambers  being  placed, 

if  necessary,  in  the  incubator 

in  tiers.     The  large  size  may 

also   be   used    in    the    same 

way  for  plate-cultivations. 
Potato       Knives.  — A 

common  broad  smooth-bladed  knife  set  in  a  wooden  handle  is  sold 

for  this  purpose. 

Scalpels.— Half  a  dozen  scalpels,  preferably  with  metal  handles, 
may  be  kept  especially  for  inoculating  sterilised  potatoes. 

Brush.— A  common  stout  nail-brush,  or  small  scrubbing-brush, 
is  essential  for  cleansing  potatoes. 


FIG.  261.— DAMP  CHAMBER  FOR  POTATO- 
CULTIVATION. 


APPARATUS,   MATERIAL,    AND   REAGENTS. 


(J)  APPARATUS  FOR  PREPARATION  OF  SOLIDIFIED  STERILE 
BLOOD-SERUM. 

Glass  Jar. — A  tall  cylindrical  glass  jar,  on  foot,  with  a  broad 

ground  stopper,  for  receiving  blood. 

Pipette. — An  ordinary  or  graduated  pipette,  for  transferring  the- 

seruin  from  the  jars  to  sterile  test-tubes  or  glass  capsules. 

Koch's     Serum     Ster- 
iliser.—A    cylindrical     case, 

with  double  walls  forming  an 

interspace  to   contain   water, 

closed  with  a  lid,  also  double- 
walled  and   provided    with  a 

tubular    prolongation    of  the 

enclosed  water-chamber  (Fig. 

262).        The     water    in    the 

cylinder  is  heated  from  below, 

and  that  in  the  lid  by  means 

of  the  prolongation. 

In     the     centre     of     the 

cylinder   is    a    column  which 

communicates  with  the  water- 

charnber  of  the  cylinder,  and 

from  it  pass  four  partitions, 

which   serve   to   support    the 

test -tubes. 

In     the     lid      are     three 

openings,  one  of  which  communicates  with  the  water-chamber  in  the 

lid  by  which  the  latter  is  filled,  and  into  which  a  thermometer  is 

then  fixed.  In  the  centre  an 
opening  admits  a  thermo- 
meter, which  passes  into  the 
central  pipe  of  the  cylinder  ; 
through  a  third  opening  a 
thermometer  passes  to  the 
cavity  of  the  cylinder.  The 
cylinder  and  cover  are  jacketed 
with  felt,  and  the  apparatus 
is  supported  on  iron  legs. 

FIG.  263. -SERUM  INSPISSATOR.  Koch's    Serum    Inspis- 

sator. — A   shallow    tin   case 
with  glass  cover,  both  case  and  cover  jjucketed  with  felt  (Fig.  263) 


FIG.  262.— KOCH'S  SERUM  STERILISER. 


<J30  APPENDICES. 

The  case  is  double-walled,  and  the  water  contained  in  the  interspace 
is  heated  from  below.  It  is  supported  on  four  legs,  and  the  two 
front  ones  move  in  grooves  in  the  case,  so  that  the  latter  can  be 
placed  obliquely  at  the  angle  required  and  secured  in  position  by 
screw-clamps.  It  is  employed  for  coagulating  sterile  liquid  serum, 
and  for  solidifying  nutrient  agar-agar  so  as  to  give  them  a  sloping 
surface. 

Hueppe's  Serum  Inspissator. — By  the  new  process  the  serum 
is  obtained  with  every  possible  precaution,  and  solidified  at  once  in 
Hueppe's  apparatus  (Fig.  44). 

Glass  Capsules. — Small  capsules  or  hollowed-out  cubes  of 
crystal  glass  are  employed  for  cultivation  on  solid  blood-serum,  on 
nutrient  gelatine,  and  on  agar-agar.  They  may  be  procured  of 
white  and  blackened  glass,  and  are  provided  with  glass  slips  as 
•covers. 


(K)  APPARATUS  FOR  STORING,  AND  FOR  CULTIVATIONS  IN,  LIQUID 

MEDIA. 

Lister's  Flasks. — Lister  devised  a  globe-shaped  flask  with  two 
necks — a  vertical  and  a  lateral  one.  The  lateral  one  is  a  bent  spout, 
tapering  towards  its  constricted  extremity.  When  the  vessel  is 
restored  to  the  erect  position  after  pouring  out  some  of  its  contents, 
a  drop  of  liquid  remains  behind  in  the  end  of  the  nozzle,  and 
prevents  the  regurgitation  of  air  through  the  spout.  A  cap  of 
cotton  wool  is  tied  over  the  orifice,  and  the  residue  in  the  flask  kept 
for  future  use.  The  vertical  neck  of  the  flask  is  plugged  with 
sterilised  cotton  wool  in  the  ordinary  way  (Fig.  60). 

Sternberg's  Bulbs. — Sternberg  advocates  the  use  of  a  glass 
bulb,  provided  with  a  slender  neck  drawn  out  to  a  fine  point  and 
hermetically  sealed  (Fig.  62). 

Aitken's  Test-tube. — This  is  an  ingenious  device  for  counter- 
acting the  danger  of  entrance  of  atmospheric  germs  on  removal  from 
the  ordinary  test-tube  of  the  cotton-wool  plug.  Each  test-tube  is 
provided  with  a  lateral  arm  tapering  to  a  fine  point,  which  is 
hermetically  sealed  (Fig.  62). 

Drop-culture  Slides.— About  a  dozen  or  more  thick  glass 
slides  with  a  circular  excavation  in  the  centre  are  required  for 
drop-cultures  (Fig.  48). 

Vaseline.— A  small  pot  of  vaseline  with  a  camel's-hair  brush 
should  be  reserved  especially  for  use  in  the  preparation  of  drop- 
cultures. 


APPARATUS,  MATERIAL,  AND  REAGENTS. 


631 


Bulbed  Tubes. — Glass  vessels,  such  as  test-tubes, 
and  pipettes,  which  are  used  in  dealing  with  liquid  media,  have 
already  been  mentioned  under  other  headings;  but  bulbed  tubes, 
Pasteur's  bulbs,  and  various  other  forms  are  also  required  for  special 
experiments. 


(L)  APPARATUS  FOR  INCUBATION. 

There  are  several  forms  of  incubator,  each  of  which  has  its 
advocates.  They  are  mostly  rectangular  chests,  with  glass  walls 
front  and  back,  or  in  front 
only.  A  cylindrical  model 
is  preferred  by  some.  Two 
only  will  be  described 
here — D'Arsonval's  and 
Babes'.  The  former  admits 
of  very  exact  regulation  of 
temperature,  and  the  latter 
is  a  very  practical  form  for 
general  use. 

D'Arsonval's  Incu- 
bator. —  The  "Etwoe. 
D'Arsonval"  (Fig.  264)  is 
a  very  efficient  apparatus, 
and  is  provided  with  a  heat- 
regulator,  which  enables 
the  temperature  to  be 
maintained  with  a  mini- 
mum variation.  It  consists 
of  a  cylindrical  copper 
vessel,  with  double  walls, 
enclosing  a  wide  interspace 
for  containing  a  large 
volume  of  water.  The  roof 
of  the  water-chamber  is 
oblique,  so  that  the  wall 

rises  higher  on  one  side  than  on  the  other.  This  admits  of  the  inter- 
space being  completely  filled  with  water.  At  the  highest  point  is 
an  opening  fitted  with  a  perforated  caoutchouc  stopper,  through 
which  a  glass  tube  passes.  The  mouth  of  the  cylinder  itself  is 
horizontal,  and  is  closed  by  a  lid,  which  is  also  double- walled  to 
contain  water.  In  the  lid  are  four  openings  :  one  serves  for  filling  its 


FIG.  264.— D'ARSONVAL'S  INCUBATOR. 


632 


APPENDICES. 


FIG.  265.— SCHLOSING'S  MEMBRANE 
REGULATOB. 


water-chamber,  and  the  others  for  thermometers  and  for  regulating 
the  air  supply  in  the  cavity  of  the  cylinder.  The  cylinder  is  con- 
tinued below  by  a  cone,  also  double- walled,  and  there  is  a  perforated 
grating  at  the  line  of  junction  of  the  cylinder  and  cone.  The  cone 
terminates  in  a  projecting  tube  provided  with  an  adjustable 
ventilator.  The  apparatus  is  fixed  on  three  supports  united  to 
one  another  below.  One  of  them  is  utilised  for  adjusting  the  height 
of  the  heating  apparatus.  Situated  above  this  leg  is  the  heat- 
regulating  apparatus  (Fig.  265),  attached  to  a  circular,  lipped 

aperture  in  the  outer  wall  of  the 
incubator.  To  the  lip  is  fixed  with 
six  screws  the  corresponding  lip  of 
a  brass  box,  with  a  tightly -stretched 
diaphragm  of  indiarubber  inter- 
vening. Thus  the  diaphragm 
separates  the  cavity  of  the  box  from 
the  water  in  the  interspace  of  -the 
incubator.  The  cap  of  the  box, 
which  screws  on,  is  bored  in  the 
centre  for  the  screw-pipe,  by  which 
the  gas  is  supplied.  Another  pipe 

entering  the  box  from  below  is  connected  with  the  gas-burners. 
Around  the  end  of  the  screw-pipe  a  collar  loosely  fits,  and  is  pressed 
against  the  diaphragm  by  means  of  a  spiral  wire  spring.  Close 
to  the  mouth  of  the  screw-pipe  a  small  opening  exists,  so  that  the 
gas  supply  to  the  burners  is  not  entirely  cut  off  even  when  the 
diaphragm  completely  occludes  the  mouth  of  the  screw-pipe. 

To  work  the  apparatus  the  tube  and  plug  must  be  removed,  and 
the  water-chamber  filled  completely  with  distilled  or  rain  water  at  the 
temperature  required.  The  caoutchouc  plug  is  replaced  and  the  tube 
placed  in  position.  Gas  enters  through  d  (Fig.  265),  and  passes  through  the 
opening  at  its  extremity  into  the  chamber  of  the  box.  Thence  it  passes 
through  the  vertical  exit  which  is  connected  with  the  gas-burners.  As 
the  temperature  rises  the  water  rises  in  the  tube,  and  at  the  same  time 
exercises  a  pressure  on  every  part  of  the  walls  of  the  incubator,  and 

icnce  on  the  diaphragm.     In  consequence  of  this,  the  diaphragm  bulging 

outwards  approaches  the  end  of  the  tube  d,.and  gradually  diminishes  the 

i  supply.     As  a  result  the  temperature  falls,  the  water  contracts  and 

sinks  m  the  tube,  and  the  diaphragm  receding  from  d,  the  gas  supply 

i  again  increased.  By  adjusting  the  position  of  the  tube  d  to  the 
diaphragm,  any  required  temperature  within  the  limits  of  the  working  of 
the  apparatus  can  be  regulated  to  the  tenth  of  a  degree- provided  (1) 
that  the  gas  supply  is  rendered  independent  of  fluctuations  of  pressure 


APPARATUS,   MATERIAL,   AND   REAGENTS. 


633 


by  means  of  a  gas-pressure  regulator  ;  (2)  that  the  height  of  the  watc  r 
in  the  tube  is  controlled  daily  by  the 
withdrawal  or  addition  of  a  few  drops  of 
distilled  water ;  and  (3)  that  the  apparatus 
is  kept  in  a  place  with  as  even  a  tempera- 
ture as  possible,  and  sheltered  from  currents 
of  air. 

The  burners  in  Fig.  264  are  protected 
with  mica  cylinders  similar  to  the  burner 
represented  in  Fig.  266.  The  flames  of 
these  burners  can  be  turned  down  to  the 
smallest  length  without  danger  of  extinction, 
and  the  temperature  may  be  regulated  very 
satisfactorily  without  using  the  heat-regulator 
just  described,  if  the  gas  first  passes  through 

a  pressure-regulator  (Fig.  269).     To  provide  FlG-   2GG.— GAS-BURNER 
against  the  danger  resulting  from  accidental 
extinction  of  the   gas,   Koch  has  devised  a 
self-acting  apparatus  (Fig.  267),  which,  simultaneously  with  the  extinction 
of  the  flame  of  the  burner,  shuts  off  the  supply  of  gas. 


FIG.  267.— KOCH'S  SAFETY  BURNER. 


Babes'  Incubator. — The  pattern  used  by  Babes  is  a  very 
simple  one,  and  may  be  recommended  for  economy  and  efficiency 
(Fig.  268). 

It  consists  of  a  double-walled  chest  with  sides  and  roof  jacketed 
with  felt.  Water  fills  the  interspace  between  the  walls,  and  on 
the  roof  are  two  apertures — one  for  a  gas-regulator  and  the  other 


634 


APPENDICES. 


for   a   thermometer.     In  front  the   chest   is   closed   in   by  a  sheet 

of  felt,  a  glass  door,  and  a  sliding 
glass  panel.  The  apparatus  can  be 
suspended  on  the  wall  or  supported 
on  legs,  and  is  heated  from  below 
by  means  of  protected  burners. 

The  gas  should  pass  first  through 
a  pressure-regulator,  and  then 
through  a  thermo-regulator  to  the 
burners. 

Moitessier's  Gas  -  pressure 
Regulator. — This  apparatus  is  best 
explained  by  reference  to  the  dia- 
gram (Fig.  269).  In  the  bottom  of 
the  cylinder  (A)  are  the  entrance  (k) 
and  exit  (I)  gas-tubes.  The  tap  (m) 
regulates  the  size  of  the  flame.  The 
cover  (n  n)  roofs  in  the  cylinder  (A). 
The  bell  (B)  supports,  by  means  of 
e  arid  /,  the  ball  valve  (d),  which 
lies  in  the  cover  (c  c).  The  gas, 
entering  by  k,  passes  through  the 

valve  (d),  and  is  thence  conducted  by  the  tube  a  to  the  tube  I.     The 

bell  (B)  and  the  weighted  dish  (h)  are  screwed  on  to  the  connecting- 
rod   (g).      To   diminish    as 

much      as      possible      the 

friction     of     g     in     i,     g 

only    touches    i    by    three 

projecting  ridges.     Section 

of  i  and  g  is  shown  at  s. 

To  put   the   apparatus   in 

use  it  is  first  levelled,  then 

h  is  screwed  off,   and   the 

cover  (n  n)  removed.      A 

mixture    of   two    parts    of 

pure  acid-free  glycerine  to 

one    of    distilled    water    is 

poured    into    the    cylinder 

until    it    flows   out    at    q, 

which  is  then  closed,  and 

the   cover   (n  n)  replaced. 

The   manometers   are    filled    with    coloured    water,    and    k    and    I 


FIG.  268. — BABES'  INCUBATOR. 


FIG.  269. — MOITESSIER'S  GAS-PRESSURE. 
REGULATOR. 


APPARATUS,   MATERIAL,    AND   REAGENTS.  635 

connected  with  the  entrance  and  exit  gas  tubing  respectively.  The 
pressure  of  the  incoming  gas  raises  the  bell  (B)  ;  and  with  it  the  valve 
(d)  is  raised  towards  the  opening  at  c  c.  The  weight  (h\  which  is 
replaced  on  #,  by  its  downward  pressure  counteracts  this  upward 
pressure  of  the  gas  and  opens  the  valve  (c  c).  Thus  the  flame  is 
best  regulated  in  the  morning,  when  the  pressure  is  at  a  minimum ; 
then  supposing  an  increase  of  pressure  occurs,  the  weight  of  h  is 
overbalanced,  B  is  raised,  and  with  it  d,  and  the  gas  supply  pro- 
portionately diminished  by  the  gradual  closing  of  the  valved  opening. 

Reichert's    Thermo-regulator.— This    regulator   (Fig.   270) 
consists   of   three   parts — a   hollow    "["-piece,    a   stem   and   a   bulb. 
The  T -piece  fits  like  a  stopper  in  the  upper  widened 
portion   of   the  stem.      One   arm  of  the  T  is  open 
and   connected   with   the   gas    supply;    the    vertical 
portion    terminates   in   a  small   orifice,    and    is   also 
provided  with  a  minute  lateral  opening.     The  stem  is 
provided  with  a  lateral  arm,  and  this  arm,  the  stem, 
and   the   bulb   contain   mercury.      The   regulator   is 
fixed  in  the  roof  of  the  incubator,   so  that  the  bulb 
projects  either  into  the  interior  of  the  incubator  or 
into  the  water-chamber.     When  the  incubator  reaches 
the  required  temperature,  the  mercury  is  forced  up 
by  means  of  the  screw  in  the  lateral  arm,  until  it      FIG.  270.— 
closes    the   orifice    at  the   extremity   of   the  vertical      RT™MO'S 
portion   of   the    T-      The  gas  which  passes  through      REGULATOR. 
the    lateral    orifice    is    sufficient    to    maintain     the 
apparatus  at  the  required  temperature.     If  the  temperature  of  the 
incubator  falls,  the  mercury  contracts,  and  gas  passing  through  the 
terminal  orifice  of  the  T  increases  the  flame  of   the  burner,  and 
the  temperature  is  restored. 

Page's  Thermo-regulator  resembles  the  above,  but  instead 
of  the  T~ -piece  there  are  two  pieces  of  glass-tubing.  The  outer 
tubing  envelops  the  upper  part  of  the  stem  of  the  regulator,  and 
admits  of  being  raised  or  lowered.  The  upper  end  of  this  tubing 
is  closed  by  a  cork,  which  is  perforated  to  admit  the  narrow  glass- 
tubing,  which  represents  the  vertical  arm  of  the  J,  passing  within 
the  stem  of  the  regulator.  This  has  a  terminal  and  a  lateral 
opening,  and  is  the  means  of  entrance  for  the  gas.  This  regulator 
is  adjusted  by  noting  when  the  thermometer  indicates  the  desired 
temperature,  and  then  pushing  down  the  outer  tube  until  the 
terminal  opening  of  the  inner  tube,  which  is  carried  down  with 
it,  is  obstructed  by  the  mercury. 


636 


APPENDICES. 


Meyer's  Thermo-regulator  is  represented  in  Fig.  271.  No.  I, 
shows  the  construction  of  the  regulator  :  its  inner  tube  terminates 
in  an  oblique  opening,  and  is  also  provided  with  a  minute  lateral 
aperture,  which  prevents  the  complete  shutting  off  of  the  gas  supply. 


FIG.  271. — MEYER'S  THERMO-EEGULATOR. 

No.  II.  illustrates  the  method  of  introducing  the  mercury  by  suction 
through  a  filling  tube,  which  is  substituted  for  the  inner  tube  of  the 
regulator.  No.  III.  represents  Frankel's  modification  of  the  same 
instrument. 


(M)  INOCULATING  AND  DISSECTING  INSTRUMENTS  AND  APPARATUS 
IN  COMMON  USE. 

Mouse-cages. — As    mice    are    the    animals    most    frequently 
employed    for    experimental     purposes,     mouse-cages     have     been 


APPARATUS,  MATERIAL,  AND  REAGENTS. 


637 


especially  introduced,  consisting  simply  of  a  cylindrical  glass  jar 
with  a  weighted  wire  cover. 

Dressing-case. — A  small  surgical  dressing-case,  with  its  usual 
accessories — forceps,  knives,  small,  straight  and  curved  scissors, 
needles,  silk,  and  so  forth — will  serve  for  most  purposes. 

Pravaz'  Syringe. — Koch's  modification  of  Pravaz'  syringe 
admits  of  sterilisation  by  exposure  to  150°  0.  for  a  couple  of  hours. 

Special  Instruments  and  Material. — Instruments  required 
for  special  operations,  and  the  materials  necessary  for  strict  anti- 
septic precautions,  need  not  be  detailed  here.* 

Dissecting-boards.— Slabs  of  wood  in  various  sizes,  or  gutta- 
percha  trays,  provided  with  large-headed  pins,  are  employed  for 
ordinary  purposes. 

Dissecting-case. — A  dissecting-case,  fitted  with  scalpels, 
scissors,  hooks,  etc.,  should  be  reserved  entirely  for  post-mortem 
examinations . 


FIG.  272.— SIPHON  BOTTLE,  WITH  FLEXIBLE  TUBE,  GLASS  NOZZLE,  AND  A 
MOHR'S  PINCHCOCK. 


(N)  GENERAL  LABORATORY  REQUISITES. 

Siphon  Apparatus. — Two  half-gallon  or  gallon  glass  bottles, 
with   siphons    connected    with    long    flexible   tubes   provided   with 
glass  nozzles  and  pinchcocks  (Fig.  272),  should  be  employed  for  the 
*    Vide  Cheyne,  Antiseptic  Sivrgery.     1882. 


638 


APPENDICES. 


following  purposes : — One  is  used  to  contain  distilled  water,  with 
the  nozzle  hanging  down  conveniently  within  reach  of  the  working 
table;  the  other  is  to  contain  a  solution  of  carbolic  acid  (1  in  20), 
and  may  be  placed  so  that  the  nozzle  hangs  close  to  the  lavatory 
sink  or  basin.  The  former  replaces  the  use  of  the  ordinary  wash- 
bottle,  in  washing  off  surplus  stain  from  cover-glasses,  etc.,  and  the 
latter  is  conveniently  placed  for  disinfection  of  vessels  and  hands 
after  cleansing  with  water.  They  should  be  placed  on  the  top  of  a 
cupboard  or  on  a  high  shelf. 

Desiccator. — The  desiccator  (Fig.  273)  consists  of  a  porcelain 

pan  containing  concentrated 
sulphuric  acid  and  covered 
over  with  a  bell-glass  receiver. 
The  sheet  of  plate-glass  upon 
which  the  pan  rests  is  ground 
upon  its  upper  surface,  and 
the  rim  of  the  glass  bell  is 
also  ground  and  well  greased. 
In  the  centre  of  the  pan  is  a 
column  supporting  a  circular 
frame,  which  is  covered  with 
wire  gauze.  Slices  of  potatoes, 
upon  which  micro-organisms 
have  been  cultivated,  are 
rapidly  dried  by  the  action  of 
sulphuric  acid  in  confined  air. 

A  detailed  description  of  other  kinds  of  apparatus  commonly  in 
use  in  a  research  laboratory — such  as  the  various  forms  of  apparatus 
for  filtering  cultures  in  liquids,  and  the  reagents  necessary  for 
special  chemical  investigations — must  be  sought  for  elsewhere. 
Much  information  may  be  obtained  about  the  most  recent  improve- 
ments in  bacteriological,  chemical  and  physical  apparatus  by 
reference  to  manufacturers'  catalogues.* 

*  All  bacteriological  apparatus  may  be  obtained  from  Berlin  from  Dr. 
Muencke,  58,  Louisen  Strasse,  or  Dr.  Hermann  Eohrbeck,  24,  Karlstrasse. 
Dr.  George  Griibler,  Leipzig,  is  recommended  for  special  staining  reagents. 
In  London,  chemicals  and  bacteriological  apparatus  can  be  obtained  from 
Becker  &  Co.,  Hatton  Wall,  or  from  Baird  &  Tatlock,  14,  Cross  Street,  Hatton 
Garden,  B.C.  Mr.  Baker,  of  High  Holborn.  W.C.,  is  recommended  as  the 
agent  for  microscopes  and  objectives  by  Continental  makers,  including 
Zeiss'  apochromatic  objectives. 


FIG.  273.  — DESICCATOE. 


APPENDIX     V. 
BIBLIOGRAPHY 


CHAPTER    I. 

HISTORICAL   INTRODUCTION. 

Andry,  De  la  Generation  des  Vers  dans  le  Corps  de  1'Homme,  1701.  Charlton 
Bastian,  Proc.  Koyal  Soc.  1872.  Bonnet,  Considerations  sur  les  Corps  orga- 
nises, 1768.  Davaine,  Compt.  Kend.,  T.  Iviii.  and  lix.  Gleichen,  Dissertation 
sur  la  Generation,  1778.  Ball,  Essays  in  Natural  History  and  Philosophy. 
Kircher,  Ars  magna  lucis  et  umbrae,  1646.  Koch,  Beitrage  zur  Biologie  der 
Pflanzen,  1876.  Lister,  Pharmaceut.  Journal  and  Transact.,  1877.  Miiller, 
Animalia  infusoria,  1786.  Pasteur,  Compt.  Kend.,  1859,  1880  ;  Etude  sur  la 
Maladie  des  Vers-a-soie,  1870.  Plenciz,  Opera  Medico-Physica,  1762.  Schroder 
and  Von  Dusch,  Ann.  der  Chem.  und  Pharm.,  vol.  Ixxxix.  Schultze,  Poggen- 
dorff's  Ann.,  vol.  xxxix.  Schwann,  Poggendorff's  Ann.,  vol.  xli.  Tyndall,  Essays 
on  floating  matter  of  the  air,  1881. 

CHAPTER   II. 

MORPHOLOGY   AND   PHYSIOLOGY   OF   BACTERIA. 

MORPHOLOGY. 

Baumgarten,  Lehrbuch  der  Pathologischen  Mykologie,  1890.  Biedert, 
Virch.  Archiv,  Bd.  100,  1885.  Billroth,  Unters.  uber  d.  Veg.  Form  der  Cocco- 
bacteria  septica,  1874.  Brefeld,  Botanische  Untersuch.  uber  Schimmelpilze, 
Heft  1,  1881.  Cohn,  Beitrage  zur  Biologie  der  Pflanzen,  Bd.  I.,  1872,  1875. 
Cornil  and  Babes,  Les  Bacteries,  1885.  Dallinger  and  Drysdale,  Monthly  Micro- 
scop.  Journ.,  1875.  De  Bary,  Verg.  Morph.  und  Biolog.  der  Pilze,  Mycetozoen, 
und  Bacterien,  1884  ;  Vorlesungen  uber  Bacterien,  1885.  Dujardin,  Histoire 
Naturelle  des  Zoophytes,  1841.  Ehrenberg,  Die  Infusionsthierchen  als  Volkom. 
Organism,  1838.  Fisch,  Biolog.  Centralbl.,  V.,  1885.  Fliigge,  Handbuch 
der  Hygiene,  1883;  Die  Micro-organismen,  2nd  Edition,  1886.  Gram, 
Fortsch  der  Med.,  II.,  No.  6.  Grove,  Synopsis  of  the  Bacteria  and  Yeast 
Fungi,  1884.  Hallier,  Die  Pflanzlichen  Parasiten,  1866.  Hauser,  Ueber 
Faulniss  Bacterien,  1885.  Hueppe,  Die  Formen  der  Bakterien,  1885.  Lankester, 
Quart.  Journ.  Microscop.  Science,  1873.  Leunis,  Synopsis  d.  Pflanzenkunde : 

639 


640  APPENDICES. 

Hanover,  1877.  Lister,  Quart.  Journ.  Microscop. '  Science,  1873.  Lutz, 
Fortschr.  d.  Med.,  1881.  Marpmann,  Die  Spaltpilze,  1884.  Miller,  Deut.  Med. 
Woch..  1884.  Nageli,  Die  Niedereu  Pilze.  1877.  Neelsen,  Biol.  Centralbl.,  III., 
No.  18,  1883.  Sachs,  Text-book  of  Botany,  1882.  Van  Tieghem,  Compt. 
Eend.,  1879 ;  Traite"  de  Botanique,  1883.  Zopf,  Die  Spaltpilze,  1885. 

GENERAL  BIOLOGY. 

Arloing,  Archiv  de  Physiol.,  1886.  Bordoni-Uffreduzzi,  Fortschr.  d.  Med., 
1886.  Cheyne,  Brit.  Med.  Journ.,  1886.  Cohn  and  Mendelsohn,  Beitr.  z. 
Biol.  d.  Pflanzen,  Bd.  III.,  Heft  1.  Cortes,  Compt.  Ptend.,  T.  99,  p.  385,  1884. 
Downes,  Proc.  Roy.  Soc.,  1886.  Duclaux,  Compt.  Rend.,  1885.  Engelmann, 
Arch.  f.  d.  Ges.  Physiologic,  Bd.  26,  1881  ;  Botan.  Zeitg.,  1882.  Fodor, 
Archiv  f.  Hygiene,  1886.  Hauser,  Archiv  f.  Exper.  Patholog.  u.  Pharmacologie, 
1886.  Hofmann,  Allgem.  Med.  Centralbl.,  S.  605.  Liborius,  Zeitschrift  f. 
Hygiene,  1886.  Nageli,  Die  Niederen  Pilze :  Miinchen,  1877  ;  Unters.  liber 
Niedere  Pilze  :  Miinchen,  1882.  Nencki,  Virchow's  Archiv,  1879  ;  Beitriige  zur 
Biol.  der  Spaltpilze:  Leipzig,  1879;  Ber.  d.  Deutschen  Chem.  Gesellsch.,  S. 
2605,  1884.  Eegnard,  Compt.  Rend.,  T.  98,  p.  744,  1884.  Tumas,  St.  Petersb. 
Med.  Wochenschr.,  1879.  Wyssokowitsch,  Zeitschrift  f.  Hygiene,  1886. 

CHROMOGENIC  BACTERIA. 

Babes,  Biolog.  Centralbl.,  Bd.  2,  1882.  Chabert  and  Fromage,  D'une  Altera- 
tion du  Lait  de  Vache,  designee  sous  le  Nom  du  Lait  Bleu.  1880.  Charrin, 
Communication  faite  a  la  Societe  Anatomique,  1884.  Cohn  mid  Miflet,  Cohn's 
Beitrage  zur  Biol.  d.  Pflanzen,  Bd.  TIL,  Heft  1.  1879.  Eberth,  Centralbl.  f.  d. 
Med.  Wissensch.,  1863.  Ehrenberg,  Micr.  Prodigiosus,  Verhandl.  d.  Berl. 
Acad.,  1839.  Fordos,  Compt.  Rend,  de  1'Acad.  de  So.,  1860.  Frank,  Cohn's 
Beitr.  z.  Biol.  d.  Pflanzen,  Bd.  I.,  Heft  3,  1875.  Gessard,  De  la  Pyocyanine 
et  de  son  Microbe,  1882  ;  Ann.  de  1'Institut  Pasteur,  T.  iv.  Gielen,  Mag.  f. 
Oes.  d.  Thierheilkunde,  1852.  Girard,  Unters.  iiber  Blauen  Eiter ;  Chirurg. 
Centralbl.,  II.,  1875;  Revue  des  So.,  T.  5,  1877.  Hermstadt,  Ueber  die 
Blaue  und  Rothe  Milch.,  1833.  Hugues,  Echo  Veterinaire,  1884.  Klein, 
Quart.  Journ.  of  Micr.  Sc.,  Vol.  15,  1875.  Lankester,  Quart.  Journ.  of  Micr.  Sc., 
Vol.  13,  1873,  1876.  Liicke,  Arch.  f.  Klin.  Chir.,  1862.  Mosler,  Virchow's 
Archiv,  Bd.  43,  1868.  Neelsen,  Cohn's  Beitr.  z.  Biologic  d.  Pflanzen,  Bd.  III., 
Heft  2,  1880.  Schrdter,  Cohn.  Beitr.  z.  Biol.  der  Pflanzen,  Bd.  I.,  Heft,  2, 
1872.  Steinhoff,  Neue  Ann.  d.  Mecklenb.  Landw.  Ges.,  1838.  Wernich,  Cohn's 
Beitrage  zur  Biol.  d.  Pflanzen,  Bd.  III.,  Heft  1,  1879.  Van  Tieghem,  Bull,  de 
la  Soc.  Bot.  de  France,  1880. 

ZYMOGENIC  BACTERIA  AND  FERMENTATION. 

Be"champ,  Compt.  Rend,,  T.  60,  p.  445,  1865  ;  T.  93,  1881.  Boutroux,  Compt. 
Rend.,  T.  86,  1878.  Brefeld,  Landwirthsch.  Jahresber.,  Bd.  3,  1874;  Bd.  4, 
1875 ;  Bd.  5,  1876.  Cienkowski,  Die  Gallertbildungen  d.  Zuckerriibensaftes, 
1878.  Colin,  Bull,  de  1'Acad.  de  Med.,  1875.  Dubrunfaut,  Compt,  Rend., 
T.  73,  1871.  Duclaux,  Theses  presentees  a  la  Faculte  de  Paris,  1865. 
Dumas,  Compt.  Rend.,  T.  75.  Nr.  6,  1872 ;  Ann.  de  Chim.  et  de  Phys.,  1874. 
Eriksson,  Unters.  aus.  d.  Botan. ;  Institut  in  Tubingen,  Heft  1,  1881.  Feltz 
and  Bitter,  Journ.  de  1'Anat.  et  Phys.,  1874.  Fermi,  Centralb.  f.  Bact., 
1891.  Fitz,  Berichte  d.  Chem.  Ges.,  Bd.  6,  S.  48,  1875;  Bd.  10,  p.  216,  1878  ; 


BIBLIOGRAPHY.  641 

Bd.  11,  pp.  42  and  498,  and  Bd.  12,  p.  474,  1879;  Bd.  13,  p.  1309,  1880; 
Bd.  15,  p.  857,  1882;  Bd.  16,  p.  844,  1883;  Bd.  17,  p.  1188,  1884.  Fleck, 
Ber.  d.  Chem.  Centralst. :  Dresden,  1876.  Frankland,  Cantor  Lectures,  1892. 
Gessard,  I)e  la  Pyocyanine  et  de  son  Microbe.  Guiard,  These  de  Paris,  1883. 
Hallier,  Gilhrungserscheinungen,  1867.  Hanson,  Untersuch.  aus.  d.  Prax.  der 
Gahrungsindustrie,  1890,  1892.  Harz,  Grundziige  der  alkoholischen  Gah- 
rungslehre,  1877.  Hiller,  Centralbl.  f.  d.  Med.  Wiss.,  S.  53,  1874.  Hofmann, 
Aerztl.  Verein  zu  Wien,  Mai  1873;  Allgem.  Med.  Centralbl.,  S.  605,  1873. 
Hoppe-Seyler,  Medic.-Chem.  Untersuchungen,  Heft  4,  1871.  Hueppe,  Mitth. 
a.  d.  Ges.  Amt,  Bd.  ii.,  1884 ;  Deut.  Med.  Woch.,  1884.  Jacksch,  Zeitschr.  f. 
Physiol.  Chemie,  Bd.  5,  1881.  Jorgensen,  Microorg.  of  Ferment  Trans.,  1893. 
Karsten,  Chemismus  der  Pflanzenzelle,  1869.  Kern,  Bull,  de  la  Soc.  Imp.  des 
Naturalistes  de  Moskau,  No.  3,  1881.  Krannhals,  Deut.  Arch.  f.  Klin.  Med., 
Bd.  35,  1884.  Ladureau,  Compt.  Kend.,  T.  99,  p.  877,  1884.  Le*pine  and  Roux, 
€ompt.  Rend.,  T.  101,  1885.  Leube,  Virch.  Arch.,  Bd.  100,  S.  540,  1885.  Lex, 
Centralbl.  f.  d.  Med.  Wiss.,  S.  291,  1872.  Liebig,  Verhandl.  der  Munchener 
Akad.  d.  Wiss.,  1861 ;  5  Nov.  1869 ;  Ueber  Gahrung,  Quelle  der  Muskel- 
kraft  und  Ernahrung ;  Leipzig  u.  Heidelberg,  1870.  Lister,  The  Pharmac. 
Journ.  and  Transact.,  1877.  Mayer,  Lehrbuch  der  Giihrungschemie ;  2  Aufl., 
1876.  Monoyer,  These  de  Strassburg,  1862.  Miiller,  Journ.  f.  Prakt.  Chem., 
1860.  Musculus,  Ber.  d.  Chem.  Ges.,  S.  124,  1874 ;  Compt.  Kend.,  T.  78,  1874. 
Hageli,  Theorie  der  Gahrung :  Munchen,  1879.  Pasteur,  Annal.  de  China,  et 
de  Phys.,  III.  Ser.,  T.  58,  1860 :  Compt.  Rend.,  1860,  1861,  1863,  1864,  1871, 
1872;  Bull,  de  la  Soc.  Chim.,  1861;  Ann.  de  Chim.  et  de  Phys.,  T.  64,  1862; 
Etudes  sur  le  Vin,  1866 ;  Bull,  de  1'Acad.  de  Med.,  No.  27,  1876  ;  Etudes  sur 
la  Biere,  1876.  Pasteur  and  Joubert,  Compt.  Rend.,  T.  83, 1876.  Popoff,  Botan. 
Jahresber.,  1875.  Prazmowski,  Untersuchungen  iiber  die  Entvvicklungs- 
geschichte  und  Fermentwirkung  einiger  Bakterien,  1880.  Richet,  Compt. 
Rend.,  T.  88,  1879.  Scheibler,  Zeitschr.  f.  Rubenzuckerindustrie,  1874. 
Schiitzenberger,  Die  Gahrungserscheinungen,  1874.  Sheridan  Lea,  Journ. 
of  Physiology,  1885.  Tre"cul,  Compt.  Rend.,  T.  61,  1865 ;  T.  65,  1867  ;  Ann, 
des  Sc.,  Ser.  7,  T.  7,  1867.  Tyndall,  Compt.  Rend.,  T.  58,  1864  ;  Essays  on 
the  Floating  Matter  of  the  Air,  1881.  Van  Tieghem,  Compt.  Rend.,  1864, 
1874,  1879,  1880,  1884. 

PHOTOGENIC  BACTERIA. 

Beyerinck,  Archiv  Neerland,  XXIII.  Fischer,  Zeitschr.  f.  .Hygiene,  1887 ; 
Centralbl.  f.  Bakteriolog.,  1888.  Forster,  Centralbl.  f.  Bakteriolog.,  1887. 
Girard,  Compt.  Rend.,  1890.  Girard  and  Billet,  Compt.  Rend.,  1889.  Katz, 
Centralbl.  f.  Bakteriol.,  1891.  Lehmann,  Centralbl.  f.  Back,  1889.  Ludwig, 
Oentralbl.  f.  Bact.,  1887. 


CHAPTER    III. 

EFFECT   OF  ANTISEPTICS   AND    DISINFECTANTS    ON   BACTERIA. 

Arloing,  Cornevin  and  Thomas,  Lyon  Med.,  1883.  Blyth,  Proc.  Roy.  Soc., 
1885.  Buchholz,  Ueber  das  Verhalten  von  Bakterien  zu  einigen  Antiseptics, 
1876 ;  Arch,  f .  Exp.  Pathol.,  Bd.  7, 1877.  Chairy,  Compt,  Rend.,  1884.  Chamber- 
land  and  Eoux,  Compt.  Rend.,  1883.  Chauveau,  Compt.  Rend.,  1883,  1884. 
Cheyne,  Antiseptic  Surgery,  1882.  Colin,  Compt.  RendM  T.  99,  18*4.  De  la 

41 


642  APPENDICES. 

Croix,  Arch.  f.  Exp.  Pathol.,  Bd.  13,  1881.  Dujardin-Beaumetz,  Bull,  de  1'Acad.. 
de  Med.  de  Paris,  1884.  Eidam,  Cobn's  Beitr.  zur  Biol.,  Bd.  I.,  Heft  3,  1875. 
Fischer,  Berl.  Klin.  Woch.,  1882.  Fischer  and  Proskauer,  Mitth.  a.  d.  Kaiserl. 
Ges.  Amt,  Bd.  II.,  1884.  Frank,  Ueber  Desinfection  von  Abtrittsgruben,  1885. 
Frisch,  Sitzungsber.  d.  Wiener  Akad.,  Bd.  75  u.  80,  1877.  Gartner  and  Plagge, 
Deut.  Med.  Woch.,  1885.  Haberkorn,  Das  Verhalten  von  Harnbakterien  gegen 
einige  Antiseptica  ;  Dissert.  Dorpat.,  1879.  Handford,  Brit.  Med.  Journ.,  1885.. 
Heydenreich,  Compt.  Rend.,  T.  98,  1884.  Hoffmann,  Experimentelle  Unter- 
suchungen  iiber  die  Wirkung  der  Ameisensaure  Diss.  Greifswald,  1884. 
Hueppe,  Mittheilg.  a.  d.  Kaiserl.  Ges.  Amt,  Bd.  I.,  S.  341,  1881  ;  Deut.  Militar- 
arztl.  Zeitschr.,  1882.  Koch,  Cohn's  Beitr.  zur  Biol.  der  Pflanzen,  Bd.  II.,. 
Heft  2,  1876  ;  Mitth.  a.  d.  Ges.  Amt,  Bd.  I.,  S.  234,  1881.  Koch  and  Gaffky, 
Arbeit,  a.  d.  K.  Gesundh.  Amt,  1885.  Koch,  Gaffky  and  Lbffler,  Mitth.  a. 
d.  Kaiserl.  Ges.  Amt,  Bd.  I.,  S.  322,  1881.  Koch  and  Wolffhiigel,  Mitth.  a.  d. 
Kaiserl.  Ges.  Amt,  Bd.  I.,  S.  301,  1881.  Kbnig,  Chirurg.  Centralbl.,  1885. 
Laillier,  Ann.  d'Hygiene,  1883.  Larrive*,  L'Eau  Oxygenee :  These  de  Paris,. 

1883.  Lassar,  Deut.  Med.  Woch.,  1880.     Lebedeff,  Arch,  de  Physiol.  Norm,  et 
Pathol.,  1882.     Maly  and  Emich,  Sitzungsber.  d.  Kais.  Akad.  d.  Wiss.  zu  Wien. 
Jan.  1883.    Marie"-Davy,  Revue  d'Hygiene,  1884.    Merke,  Virchow's  Archiv,. 
Bd.    81,   1880.     Meyer,   Ueb.  d.  Milchsaureferment   u.    sein  Verhalten  gegen 
Antiseptica,  1880.     Mignet,  Annuaire  de  1'Observatoire   de  Montsouris,  1884- 
Miquel,  Semaine  Meclicale,  1883.     Mdrscheli,  Deut.  Med.  Woch.,  1880.     Nageli, 
Die  Niederen  Pilze,  1877.     Pasteur,  Ann.  d'Hyg.,  1880  ;  La  Vaccination  Char- 
bonneuse,  1883.     Perroncito,  Arch.  Ital.  de  Biol.,  1883.     Pictet  and  Young, 
Compt.  Rend.,  T.  98,  1884.     Plaut,  Desinfection  der  Viehstalle,  1884.     Eeinl, 
Prager  Med.  Woch.,  Nr.  10  u.  11, 1885.     Eochefort,  Herscher,  Revue  d'Hygiene, 

1884.  Eossbach,  Berl.  Klin.  Woch.,  1884.     Schede,  Sammlung  Klin.  Vortrage, 
Nr.  25,  1885.     Schill  and  Fischer,  Mitth.  a.  d.  Kaiserl.  Ges.  Amt,  Bd.  II.  1884. 
Schnetzler,  Archiv  de  Geneve,  1884.     Schrbter,    Cohn's  Beitr.  zur.  Biol.  der 
Pflanzen,  Bd.  I.,  Heft  3,  1875.     Schultz,    Deut,  Med.   Woch.,   Nr.   17,  1883 ; 
Nr.  24,  1885.     Schwartz,  Sitzungsber.  d.  Dorpater  Naturf.  Ges.,  1879.     Soyka, 
Ber.  d.  Bayr.  Akad.  d.  Wissensch.,  1879.    Steinmeyer,  Ueber  Desinfection slehre, 
1884.    Sternberg,  Amer.  Journ.  Med.  Soc.,  1883  ;    Report  of  Com.  on  Disin- 
fectants,  1888.    Thol,  Ueber  d.   Einfluss  nicht  aromat.    organ.  Sauren.    auf 
Faulniss  u.   Gahrung :    Diss.  Greifswald,   1885.     Toussaint,  Bull,   de  1'Acad., 
1880.    Tyndall,  Phil.  Trans,  of  the  Roy.  Soc.,  1877.    Vallin,  Ann.  d'Hyg.,  1877; 
Traite  des  Disinfectants  et  de  la  Desinfection,  1883 ;  Les  Nouvelles  Etuves 
&  Desinfection  :   Revue  d'Hygiene,  1883 ;   Ann.    d'Hygiene,    1884.  Wernicke, 
Virchow's  Archiv,  Bd.  78,  1879 ;  Diss.  Dorpat.,  1879.     Grundriss  der  Desin- 
fectionslehre,  1880.    Wolff,  Centralbl.  f.  d.  Med.  Wiss.,  Nr.  11,  1885.    Wolff- 
hiigel, Mittheilg.  a.  d.  Kais.  Ges.  Amt.,  Bd.  I.,  S.  188,  1881.     Wolffhiigel  and 
Knorre,  Mitth.  a.  d.  Kaiserl.  Ges.  Amt,  Bd.  I.,  S.  352,  1881. 


CHAPTER    IV. 

CHEMICAL   PRODUCTS   OF   BACTERIA. 

Backlisch,  Ber.  d.  Deutsch.  Chein.  Gesellsch.,  Bd.  18,  1880.  Bergmann, 
Das  Putride  Gift.,  1866 ;  Deut.  Zeitschr.  f.  Chirurgie,  Bd.  I.,  1872.  Bergmann 
and  Angerer,  Wiirzburger  Jubil.  Festschr.,  1882.  Bergmann  and  Schmiedeberg, 
Med.  Centralbl.,  1868.  Blumberg,  Virch.  Arch.,  Bd.  100,  S.  377,  1885.  Boeci, 


BIBLIOGRAPHY.  643 

Centralbl.  f.  d.  Med.  Wiss.,  1882.  Bouchard,  Compt.  Rend,  de  Biol.,  1882. 
Brieger,  Zeitschr.  f.  physiol.  Chemie,  Bd.  7,  1883 ;  Ber.  d.  Deutsch.  Chem.  Ges.,, 
Bd.  17,  1884  ;  Berl.  Klin.  Woch.,  Nr.  14,  1884  ;  Ueber  Ptomaine,  1885;  Weitere 
Untersuchungen  ttber  Ptomaine,  1885;  Ueber  Ptomaine:  Berl.  Klin.  Woch., 
1886.  Brouardel  and  Boutmy,  Compt.  Rend.,  T.  92,  p.  1056;  1881.  dementi 
and  Thin,  Wien.  Med.  Jahrb.,  1873.  Eber,  Centralb.  f.  Bact.,  1892.  Etard  and 
Olivier,  Compt.  Rend.,  1882.  Frisch,  Exper.  Studien  iib.  d.  Verbreitung  d. 
Fiiulnissorganismen,  1874.  Gautier,  Compt.  Rend.,  T.  94,  1882.  Gautier  and 
Etard,  Compt.  Rend.,  T.  94,  1882.  Groebner,  Beitrage  z.  Kenntniss  der 
Ptomaine,  1882.  Guareschi  and  Mosso,  Arch.  Ital.  de  Biolog.,  1883.  Hauserr 
Ueber  Faulnissbakterien,  1885.  Hemmer,  Exper.  Studien  iiber  d.  Wirkung 
Faulender  Stoffe,  1866.  Hiller,  Centralbl.  f .  Chirurgie,  1876  ;  Die  Lehre  von  der 
Faulniss,  1879.  Husemann,  Arch.  d.  Pharmac.,  1880,  1882,  1883.  Kaufmann, 
Journ.  f.  Prakt.  Chemie,  Bd.  17,  1878.  Kehrer,  Archiv  f.  Exper.  Pathol.r 
Bd.  I.,  1874.  Konig,  Ber.  iib  d.  Veterinarwesen  im  Konigreich  Sachsen. 
1881.  Maas,  Fortschr.  d.  Med.,  II.,  729,  1884.  Martin,  Rep.  Med.  Off.  Local 
Govt.  Board,  1890-91.  Nencki,  Ueber  die  Zersetzung  der  Gelatine  und  des 
Eiweisses  bei  der  Faulniss  mit  Pancreas,  1876;  Journ.  f.  Pract.  Chem., 
Bd.  26,  1882.  Offinger,  Die  Ptomaine,  1885.  Otto,  Anleitung  zur  Ausmittelung 
der  Gifte,  1884.  Panum,  Virchow's  Arch.,  Bd.  60,  1874.  Eaison,  Zur  Kenntniss 
der  Putriden  Intoxication,  1866.  Eavitsch,  Zur  Lehre  von  der  Putriden  In- 
fection, 1872.  Eosenbach,  Deut.  Zeitschrift  fiir  Chir.,  XVI.,  S.  342,  1882. 
Salomonsen,  Die  Faulniss  des  Blutes,  1877.  Schiffer,  Arch.  f.  Anat.  u.  Physiol. : 
Physiol.  Abtheil,  1882.  Schweninger,  Ueber  d.  Wirkung  Faulender  Org.  Sub- 
stanzen,  1866.  Selmi,  Chemische  Ber.,  Bd.  6,  7,  12,  1878.  Tanret,  Compt. 
Rend.,  T.  92,  1881.  Tappeiner,  Med.  Centralbl.,  1885.  Vandevelde,  Arch,  de 
Biol.  par  van  Beneden,  1884.  Willgerodt,  Ueber  Ptomaine,  1884.  Zulzer  and 
Sonnenschein,  Berl.  Klin.  Woch.,  1869. 


CHAPTER  Y. 

IMMUNITY. 

Arloing,  Cornevin  and  Thomas,  Du  Charbon  Bacterien;  Pathogenic  et 
Inoculations  Preventives,  1883.  Behring,  Deutsche  Med.  Woch.,  1890. 
Behring  and  Kitasato,  Deutsche  Med.  Woch.,  1890.  Blazekovic,  Oesterr 
Monatschr.  f.  Thierheilk.,  1884.  Bouchard,  Compt.  Rend.,  1889.  Bouley, 
L'Inoculation  Preventive  de  la  Fievre  Jaune :  Compt.  Rend.,  T.  100,  1885. 
Brieger  and  Frankel,  Berl.  Klin.  Woch.,  1890.  Buchner,  Eine  neue  Theorie 
iiber  Erzielung  v.  Immunitat  gegen  Infectiouskrankheiten,  1883.  Chamberland, 
Le  Charbon  et  la  Vaccination  Charbonneuse  d'apres  les  Travaux  R6cents  de 
M.  Pasteur,  1883.  Chamberland  and  Eoux,  Compt.  Rend.,  T.  96,  Nr.  15,  1883. 
Chauveau,  Compt.  Rend.,  T.  89,  1879;  T.  96,  Nr.  9;  Nr.  10;  Nr.  11,  1883;  Gaz. 
Hebdom.  de  Med.  et  de  Chir.,  22,  1884.  Feltz,  Compt.  Rend.,  T.  99,  p  246, 
1884.  Frank,  Jahresber.  d.  K.  Thierarzneischule  in  Munchen,  1883.  Gamaleia, 
La  Semaine  Med.,  1890.  Grawitz,  Die  Theorie  der  Schutzimpfung  :  Virchow's 
Arch.,  Bd.  48,  1881.  Hankin,  Brit.  Med.  Journ.,  1889  and  1890;  Proc.  Roy. 
Soc.,  1890;  Centralb.  f.  Bacteriolog.,  Bd.  IX.,  Lancet,  1891.  Hess,  Schweiz. 
Arch.  f.  Thierheilk.,  Bd.,  27, 1885.  Kitasato,  Zeitschr.  f.  Hygiene,  Bd.  X.  Koch, 
Ueber  die  Milzbrandimpfung,  1882.  Koch,  Gaff ky  and  Loffler,  Mitth.  a.  d.  Ges. 
Amt,  Bd.  II.,  1884.  Loffler,  Mitth.  a.  d.  Ges.  Amt,  Bd.  I.,  1881.  Martin,  Reports 


APPENDICES. 

Med.  Dept.  Loc.  Govt.  Board,  1890-91 ;  Brit.  Med.  Journal,  1891.  Masse*,  Des 
Inoculations  Preventives  dans  les  Maladies  Virulentes,  1883.  MetschnikofF, 
Virchow's  Archiv  XCVI.  and  XCVIL,  Ann.  de  1'Institut  Pasteur,  1887,  1889, 

1890,  1891,  1895.    Nuttall,  Zeitschr.  f.  Hygiene,  1888.     Oemler,  Arch.  f.  Wiss. 
u.  Pract.  Thierheilk.,  1876, 1881.    Ogata,  Centralb.  f.  Bacteriolog.,  1891.    Ollive, 
Compt.  Rend.,  T.  89,   1879.      Pasteur,   Bull,  de  1'Acad.   de   Med.   and   Gaz. 
Med.  de  Paris,  Nr.  18,  1880;  Corapt.  Kend.,  1883;  La  Vaccination  Charbon- 
neuse,   1883;  'Revue   Scientifique,    1883;    Bull,   de  1'Acad.   de    Med.,    1883. 
Perroncito,    Atti   R.   Ace.   d.    Lincei.,    1883.     Piitz,    Vortrage    f.   Thierarzte, 
Ser.  7,  Heft  1,  1884.     Roux,  Ann.  de  1'Institut  Pasteur,  1888.      Roszahegyi, 
Pester  Med.-Chir.  Presse,  1882.     Salmon  and  Smith,  Centralb.  f.  Bacteriolog., 
1887.    Semmer,   Virchow's  Arch.,    Bd.    83,   1881.     Semmer    and    Krajewski, 
Centralbl.  f.  d.  Med.  Wiss.,  1880.     Strebel,  Schweiz.  Arch.  f.  Thierheilk.,  1885. 
Tizzoni  and   Cattani,   Centralb.   f.   Bacteriolog.,  1891.     Toussaint,   Bull,    de 
1'Acad.  de  Med. ;  and  Compt.  Rend.,  1880,  1881  ;  Gazette  Medicale  de  Paris, 
Nr.  32,  1881.     Wooldridge,  Proc.  Roy.  Soc.,  1887  ;  Archiv  f.  Anat.  and  Phys., 
1888. 

CHAPTER    VI. 

ANTITOXINS   AND   SERUM-THERAPY. 

Be*clere,  Chambon  and  Me"nard,  Ann.  de  1'Institut  Pasteur,  1896.     Behring 
and  Kitasato,  Deutsche  Med.  Woch.,  1890  ;  Trans.  Internat.  Cong.  f.  Hygiene, 

1891.  Behring  and  Wernicke,  Zeitschrift  f.  Hygiene,  1892.     Buchner,  Munich 
Med.  Woch.,  1891.     Calmette,  Ann.  de  1'Institut  Pasteur,  1895.     Emmerich  and 
Mastbaum,  Archiv  f.  Hygiene,  1891.     Fedoroff,    Zeitschr.   f.   Hygiene,   1893. 
Gromakowsky,  Ann.  de  1'Institut  Pasteur,  1895.      Heubner,  Trans.  Internat. 
Med.   Congress,    1894.      Hewlett,   Practitioner,    1895.     Kossel,    Zeitschrift  f. 
Hygiene,  1894.     Marchoux,  Ann,  de  1'Institut  Pasteur,  1895.     Marmorek,  Ann. 
de  1'Institut   Pasteur;  1895,    1896.      Ogata,    Centralb.   f.   Bakteriolog.,   1891. 
Report,  Med.  Sup.  Metropolitan  Asylums  Board,  1896.     Roux,  Trans.  Internat. 
Congress  of  Hygiene,  1894 ;  Ann.  de  1'Institut  Pasteur,  1894.    Roux,  Martin, 
Chaillou,  Ann.  de  1'Instit.  Pasteur,  1894.     Tizzoni  and  Cattani,  Centralb.  f. 
Bakteriolog.,  1891.    Welch,  Trans.  Assoc.  Amer.  Physicians,  1895.    Wladmoroff, 
Zeitschr.  f.  Hygiene,  1893. 

CHAPTERS    VII.,    VIII.,    IX.,    X. 

VII. — THE  BACTERIOLOGICAL  MICROSCOPE.  VIII. — MICROSCOPICAL 
EXAMINATION  OF  BACTERIA.  IX. — PREPARATION  OF  NUTRIENT 
MEDIA  AND  METHODS  OF  CULTIVATION.  X. — EXPERIMENTS 
UPON  THE  LIVING  ANIMAL. 

Almquist,   Hygeia,  XLV. ;  Stockholm,  1883.     Banti,  Manuale   di   Tecnica 
Batteriologica,  1885.   Baumgarten,  Zeitschr.  f.  Wissensch.  Mikr.,  1884.   Behrens, 
llfsbuch  zur  Ausfuhrung  Mikroskop.  Untersuch.,  1883.    Bizzozero  and  Firket, 
Manuel  de  Microscopie  Clinique,  1885.     Blanchard,  Rev.  Inter.  Sci.,  III.,  1879. 
Bordoni    Uffreduzzi,   Microparasitici,    1885.      Brefeld,    Bot.    Untersuch.  iiber 


Schimmelpilze,  Bd.  IV.,  1881 ;  Botan.  Unters.  iiber  Hef enpilze,  Bd.  V.,  1883 ; 
sik.  Med.   Ges..  in  Wurzburg,   1875.      Buchner,  In  Nageli's 


Verhandl.  d.  Physik. 


BIBLIOGRAPHY.  645 

Untersuch.  iiber  Niedere  Pilze:  Munich,  1882;  Aerztl.  Intelligenzbl.,  No.  33, 

1884.  Carpenter,  The  Microscope  and   its   Revelations  (6th   Edition),  1881. 
Cohn,  Beitrage  zur  Biologic  der  Pflanzen,  Bd.  I.,  Heft  3,  1875,  1876.     Corni! 
and  Babes,  Les  Bacteries,  188(5.     Crookshank,  Manuel  Pratique  de  Bacterio- 
logie,  traduit  par  Bergeaud ;  avec  4  Photomicrographies,  1886.     Dolley,  Tech- 
nology of  Bacteria  Investigation,  1885.     Duclaux,  Ferments  et  Maladies,  1881. 
Ehrlich,  Deut.  Med.  Woch.,  No.  19,  1882  ;  Zeitschr.  f.  Klin.  Med. ;  Bd.  I.  1880. 
Bd.  IL,  Heft  3,  1881.    Eisenberg,  Bakteriologische  Diagnostik,  1886.    Esmarch, 
Zeitschrift  f.  Hygiene,  1886.     Fehleisen,  Ueber  Neue  Method,  der  Untersuch. 
u.  Cultur  Pathogen.  Bakterien ;  Physik.  Med.  Ges.  zu  Wiirzburg,  1882.    Fliigge, 
Handbuch  der  Hygiene  und  der   Gewerbe  Krankheiten,   1883.     Friedlander, 
Microscopische  Technik  (1st  Edition),  1884.     Gibbes,  Practical  Histology  and 
Pathology,  1885.     Gram,  Fortsch.  d.  Med.,  II.,  No.  6,  1884.     Kauser,  Ueber 
Faulnissbacterien ;  mit  15  Tafeln  in  Lichtdruck,  1885.     Hazlewood,  American 
Monthly  Microscop.  Journ.,  1883.     Hiiber  and  Broker,  Die  Path.  Histolog.  und 
Bacteriologischen    Untersuch.    Methoden,    1886.      Hueppe,    Bakteriologische 
Apparate  :  Deut.  Med.  Woch.,  1886 ;  Die  Methoden  der  Bakterien  Forschung, 
1886 ;  translated  by  Biggs,  1886.     Johne,  Ueber  die  Kochschen  Reinculturen, 

1885.  Klebs,  Archiv  f.  Exp.  Pathoi.,  Bd.  I.,  1873.     Klein,  Micro-organisms  and 
Disease,  1886.     Koch,  Biol.  Klin.  Woch.,  No.  15,  1882  ;  Mitth.  a.  d.  Kais.  Ges. 
Amt,   Bd.   I.,    1881.;    Bd.   II.,   1884;    Untersuchungen   iiber  Wundinfections 
Krankheiten,  1878;  Beitrage  z.  Biol.  d.  Pflanzen/  Bd.  II.,  Heft  3,  1877.     Lee, 
The  Microtomist's  Vade  Mecum,  1885.     Magnin  and  Sternberg,  Bacteria,  1884. 
Malley,  Photomicrography,  1885.    Orth,  Path.  Anat.  Diagnostik,  1884.    Pasteur, 
Etudes    sur  la  Biere,    1867.     Perty,    Zur   Kenntniss    Kleinster  Lebensform, 
1852.     Plant,  Farbungs  Methode  z.  Nachweis.  der  Micro-organismen,  1885 ; 
Salomonsen,  Bot.  Zeit.,  No.  39,  1879  ;  No.  28,  1880.    Schafer,  Course  of  Practical 
Histology,  1877.    Woodhead  and  Hare,  Pathological  Mycology,  1885. 


CHAPTER   XL 

EXAMINATION   OF   AIR,    SOIL   AND   WATER. 

Angus  Smith,  Rep.  to  the  Loc.  Gov.  Board,  1884 ;  Sanitary  Record,  1883. 
Becker,  Reichsmedicinalkalender,  1885.  Beumer,  Deut.  Med.  Woch.,  1886. 
Bischof,  Journ.  Soc.  Chem.  Industry,  1886.  Biichner,  Vortrage  im  Aerztl. 
Verein  zu  Munchen,  1881.  Chamberland,  Compt,  Rend.,  T.  99,  p.  247,  1886. 
Cramer,  Die  Wasserversorgung  von  Zurich,  1885.  Crookshank,  Notes  from  a 
Bact.  Labor.,  Lancet,  1885.  Cunningham,  Micr.  Exam,  of  the  Air :  Calcutta, 
1874.  Fodor,  Hygienische  Unters.  iiber  Luft,  Boden  u.  Wasser.,  1882. 
C.  Frankel,  Zeitschr.  f.  Hygiene,  1886.  Frankland,  P.  and  G.,  Proc.  Roy.  Soc., 
1885,  1886 ;  Microorganisms  in  Water,  1894.  Gunning,  Arch.  f.  Hyg.,  3,  1883. 
Heraeus,  Zeitschr.  f.  Hygiene,  1886.  Hesse,  Deut.  Med.  Woch.,  Nr.  51,  1884 ; 
2,  1884 ;  Mitth.  a.  d.  Ges.  Amt,  Bd.  II.,  1884 ;  Ueber  Wasserfiltration  :  Deut. 
Med.  Woch.,  1885 ;  Zeitschr.  f.  Hygiene,  1886.  Klebs  and  Tommasi-Crudeli, 
Archiv  f.  Exper.  Path.,  Bd.  11,  1879.  Koch,  Mitth.  a.  d.  Ges.  Amt,  Bd.  I., 
1881.  Laurent,  Journal  de  Pharmacie  et  de  Chimie,  1885.  Lemaire,  Compt. 
Rend.,  T.  57,  1863.  Letzerich,  Exp.  Unters.  iib.  die  Aetiologie  des  Typhus 
mit  bes.  Beriicksichtigung  der  Trink.  u.  Gebrauchswasser,  1883.  Maddoz, 
Month.  Microscop.  Journal,  1870.  Meade  Bolton,  Zeitschr.  f.  Hygiene,  1886. 
Miflet,  Cohn's  Beitr.  z.  Biol.  d.  Pflanzen,  Bd.  III.,  1879.  Miquel,  Annuaire 


646  APPENDICES. 

de  1'Observat.  de  Montsouris,  1877,  1882  ;  Compt.  Rend.,  T.  86,  1878  ;  Bull, 
de  la  Soc.  Chim.,  1878;  Ann.  d'Hygiene,  1879;  Les  Organismes  Vivants 
de  1' Atmosphere,  1883.  Miquel  and  Frendenreich,  La  Semaine  Medicale,  1884. 
Moreau  and  Planty mansion,  La  Semaine  Medicale,  1884.  Nageli,  Unters.  iiber 
Niedere  Pilze.,  1882.  Olivier,  Les  Germes  de  1'Air,  These,  Rev.  Scientif.. 
1883.  Pasteur,  Ann.  de  Chim.  et  de  Phys.,  T.  64, 1862 ;  Compt.  Rend.,  T.  50, 
1860;  T.  52,  1861;  T.  56,  1863;  T.  85,  1877.  Pfeiffer,  Zeitschr.  f.  Hygiene, 
1886.  Poachet,  Compt.  Rend.,  T.  47,  1858.  Schrakamp,  Archiv  f.  Hygiene, 
Bd.  II.,  1884.  Sehlen,  Fortschr.  d.  Med.,  Bd.  II.,  S.  585,  1885.  Smart,  Germs, 
Dust  and  Disease,  1883.  Soyka,  Sitz.-Ber.  der.  K.  Bayr.  Akad.  d.  Wiss.  :  Math. 
Physik.  Classe,  1881  ;  Vortrage  im  Aerztl.,  Vereiri  in  Miinchen,  1881;  D. 
Vierteljsch.  f.  Oeff.  Ges.,  Bd.  14,  1882;  Prager  Med.  Woch.,  1885;  Fortschr. 
d.  Med.,  1885.  Tissandier,  Compt.  Rend.,  T.  78,  1874.  Torelli,  La  Malaria 
in  Italia,  1883.  Tyndall,  Brit.  Med.  Journ.,  1877;  Essays  on  the  Floating 
Matter  of  the  Air,  1881 ;  Med.  Tim.  and  Gaz.,  1870.  Wernich,  Cohn's  Beitrage 
z.  Biol.  d.  Pflanzen,  Bd.  III.,  S.  105,  1879.  Wolffhiigel  and  Eiedel,  Arbeit. 
a.  d.  K.  Ges.  Amt,  1886.  Wollny,  Viert.  f.  Oeff.  Ges.,  S.  705, 1883.  Zander, 
Centralbl.  f.  Allg.  Ges.,  1883. 


CHAPTER    XII. 

PHOTOGRAPHY   OF   BACTERIA. 

Crookshank,  Photography  of  Bacteria,  1887.  Frankel  and  Pfeiffer,  Mikro- 
photog.  Atlas,  1889.  Giinther,  Photogram.  Path.  Mikroorg.,  1887.  Itzerott 
and  Niemann,  Atlas  der  Microphotograph,  1895.  Koch,  Cohn's  Beitrage  zur 
Biol.  der  Pflanz.,  1877  ;  Mitth.  a.  d.  K.  Gesundheitsamte,  Bd.  I.,  1881.  Neuhaus, 
Centralbl.  f .  Bakteriolog.,  Bd.  IV.  Sternberg,  Photomicrographs  and  How  to 
Make  them,  1884.  Woodward,  Rep.  to  Surg.  Gen.  U.  S.  Army,  1870. 


CHAPTER    XIII. 

SUPPURATION.     PY^MIA.     SEPTICAEMIA.     ERYSIPELAS.     GONORRHCEA. 

OPHTHALMIA. 

Ainstie,  Lancet,  1870.  Arloing,  Recherches  sur  les  Septicemies,  1884. 
Babes,  Compt.  Rend.,  1883.  Balfour,  Edinb.  Med.  Journ.,  1877.  Barthold, 
Pyaemisch-Metast.  Dissert.  Berlin,  1875.  Bastian,  Brit.  Med.  Journ.,  1878. 
BSchamp,  Compt.  Rend.,  1881;  Trans.  Internat.  Med.  Cong.  London,  1881. 
Beck,  Rep.  LOG.  Govt.  Board,  1880.  Birch-Hirschfeld,  Untersuchungen  iiber 
Pyamie,  1873.  Braidwood  and  Vacher,  Brit.  Med.  Journ.,  1882.  Burdon- 
Sanderson,  Trans.  Path.  Soc.,  1872;  Brit.  Med.  Journ.,  1875.  Crookshank, 
Trans.  Internat.  Congr.  of  Hygiene,  1892.  Dowdeswell,  Quart.  Journ.  Micr. 
Sc.  London,  Vol.  18,  1878 ;  Proc.  Roy.  Soc.  London,  Vol.  34,  1883.  Dreschfeld, 
Brit.  Med.  Journ.,  1883.  Drysdale,  Pyrexia,  1880.  (Jarre*,  Fortschritte  d. 
Med.,  165,  1885.  Heiberg,  Die  Puerperalen  u.  Pyamischen  Processe,  1873. 
Hoffa,  Fortsch.  d.  Med.,  1885.  Horsley,  Rep.  Med.  Officer  Loc.  Govt.  Board, 
1881.  Klemperer,  Zeitschr.  f.  Klin.  Med.,  1885.  Koch,  Wundinfectionskrank- 
heiten:  Leipzig,  1878;  Mittheil.  d.  Kais.  Ges.  Amts,  Bd.  I.,  1881.  Lister, 
Lancet,  1867;  Med.  Times  and  Gazette,  1877;  Quart.  Journ.  Microscop.  Science, 


BIBLIOGRAPHY.  647 

1887.  Oertel,  Zur  Aetiologie  der  Infectionskrankheiten,  1881.  Ogston,  Brit. 
Med.  Journ.,  Vol.  I.,  1881  ;  Journ.  of  Anat.  and  Phys.,  Vol.  17,  1882.  Passet, 
Ueber  Mikroorganismen  der  Eitrigen  Zellgewebsentziindung  des  Menschen  ; 
Fortschritte  d.  Med.,  Nr.  2,  1885,  Bd.  3,  1885.  Ferret,  De  la  Septicemie  : 
Paris,  1880.  Eindfleisch,  Lehrb.  der  Pathol.  Gewebelehre :  1  Aufl.,  S.  204, 
1866.  Rosenbach,  Mikrooganismen  bei  den  Wundinfectionskrankheiten  des 
Menschen :  Wiesbaden,  1884.  Steinberg,  Amer.  Journ.  Med.  Sc. ;  Johns 
Hopkins  Univ.  Stud.  Biol.  Lab.,  1882.  Steven,  Glasgow  Med.  Journal,  1884. 
Sutton,  Trans.  Path.  Soc.,  1883.  Tiegel,  Ueber  d.  Fiebererregenden  Eigen- 
schaften  des  Microsporon  Septicum :  Bern.  Diss.,  1871 ;  Virchow's  Archiv,  Bd. 
60,  1874.  Waldeyer,  Virchow's  Arch.,  Bd.  40,  1867  ;  Vortrag.  i.  d.  Med  Ges. 
zu  Breslau,  1871.  Watson-Cheyne,  Trans.  Path.  Soc.,  xxxv.,  1884. 

OSTEOMYELITIS. 

Becker,  Deut.  Med.  Woch.,  and  Berl.  Klin.  Woch.,  1883.  Collmann,  Bak- 
terien  im  Organismus  eines  an  einer  Verletzung  am  Oberschenkel  verstorbenen 
Madchens:  Gottingen,  1873.  Colzi,  Lo  Sperimentale,  1890.  Courmont  and 
Jaboulay,  Compt.  Rend.  Soc.  de  Biolog.,  1890.  Eberth,  Virchow's  Arch., 
Bd.  65,  1875.  Fehleisen,  Phys.  Med.  Ges.  Wiirzburg,  1882.  Friedmann,  Berl. 
Klin.  Woch.,  18^6.  Garre,  Fortschr.  d.  Med.,  1885.  Giordano,  Prog.  I.  Mic. 
Pyog.  infett.  u.  Eziolog.  d.  Osteom.  Impett.  Acuta,  1888.  Kranse,  Fortschr.  d. 
Med.,  Bd.  2,  1884.  Lannelongue  and  Achard,  La  Semaine  Med.,  1800 ;  and 
Compt.  Rend.  Soc.  cle  Biolog.,  1890.  Peyroud,  Compt.  Rend.,  1884.  Eodet, 
Compt.  Rendus,  T.  99,  1884.  Rosenbach,  Centralbl.  f.  Chirurgie,  1884. 
Schiiller,  Centralbl.  f.  Chirurgie,  Nr.  12,  1876. 

ENDOCARDITIS. 

Birch-Hirschfeld  and  Gerber,  Archiv  d.  Heilkunde,  1876.  Bramwell, 
Diseases  of  the  Heart,  1884.  Bristowe,  Brit.  Med.  Journ.,  1884.  Coupland, 
Brit.  Med.  Journ.,  1885.  Gibbes,  Brit.  Med.  Journ.,  1884.  Goodhart,  Trans. 
Path.  Soc.,  vol.  xxxi.,  1880.  Hamburg,  Berlin:  Inaug.-Diss.,  1880.  Zlebs, 
Archiv  f.  Exper.  Pathol.,  Bd.  9,  1878.  Koch,  Mittheil.  a.  d.  Kais.  Ges.  Amt, 
Bd.  I.,  1881.  Koester,  Virchow's  Arch.,  Bd.  72,  1875.  Kundrat,  Sitz.-Ber. 
d.  Kais.  Acad.  d.  Wissensch.  zu  Wien,  1883.  Leyden.  Zeitschr.  f.  Klin.  Med., 
1881.  Nocard,  Recueil  de  Med.  Vet.,  1885.  Oberbeck,  Casuistische  Beitrage 
zur  Lehre  von  der  Endocarditis  Ulcerosa  :  Inaug.-Diss.,  1881.  Orth,  J.,  Ver- 
sammlung  Deutscher  Naturf.  zu  Strasburg,  1885.  Osier,  Brit.  Med.  Journ.,  1885 ; 
Trans.  Int.  Med.  Congress,  1881.  Ribbert,  Fortsch.  d.  Med.,  1886.  Rosenbach, 
Archiv  fiir  Exper.  Pathol.,  Bd.  9,  1878.  Wedel,  Berl.  Klin.  Wochenschr.,  1877. 
Weichselbaum,  Wien.  Med.  Woch.,  1885.  Weigert,  Virchow's  Arch..  Bd.  84, 
1881.  Wilks,  Brit.  Med.  Journ.,  1882.  Wyssokowitsch,  Centralbl.  f.  d.  Med. 
Wissensch.,  Nr.  33,  1885. 

ERYSIPELAS. 

Baader,  Schweiz.  Naturf.  Gesellsch.,  1875.  Dennce,  Etude  sur  la  Pathogenic 
et  1' Anatomic  Pathologique  de  1'Erysipele,  1885.  Dupeyrat,  Recherches 
Cliniques  et  Experimentales  sur  la  Pathogenic  de  1'Erysipele,  1881.  Fehleisen, 
Wurzburger  Phys.  Med.  Ges.,  1881;  Deut.  Zeitschr.  f.  Chir.,  Bd.  16,  1882;  Die 
Aetiologie  des  Erysipals. :  Berlin,  1883.  Httter,  Med.  Centralbl.,  Nr.  34,  1868. 


648  APPENDICES. 

Janicke  and  Neisser,  Centralbl.  f.  Chir.,  Nr.  25,  1884.  Klebs,  Archiv  f.  Exper. 
Pathol.  u.  Pharmacol.,  Bd.  4,  1875.  Lukomsky,  Virchow's  Archiv,  Bd.  60,  1874, 
Nepven,  Des  Bacteries  dans  1'Erysipele,  1885.  Orth,  Archiv  f.  Exper.  PathoL 
u.  Pharmacol.,  Bd.  I.,  1873.  Eaynaud,  Union  Med.,  1873,  Eecklinghausen  and 
Lankowski,  Virchow's  Arch.,  Bd.  60,  1874.  Eheiner,  Virchow's  Arch.,  Bd.  100r 
Heft  2,  1884.  Tillmanns,  Verhandl.  d.  Deutsch.  Ges.  f.  Chirurgie,  1878; 
Archiv  f.  Klin.  Chirurgie,  Bd.  23,  1879.  Trosier,  Bull.  Soc.  Anat.  de  Paris, 
1875.  Wolff;  Virchow's  Arch.,  Bd.  81,  1880. 

PUERPERAL  FEVER. 

Aufrecht,  Naturforsch.  Versamml.,  1881.  Dole*ris,  La  Fievre  Puerperale  et 
les  Organismes  Infect.,  1886.  Heiberg,  Die  Puerperalen  und  Pyamischen 
Processe,  1873.  Karewski,  Zeitschr.  f.  Geburtsh.  u.  Gynakologie,  Bd.  7,  1881. 
Laffter,  Bresl.  Aerztl.  Ztg.,  1879.  Mayrhofer,  Monatsschr.  f.  Geburtsk.  u. 
Frauenkrankheiten.  Bd.  25,  1865.  Orth,  Virchow's  Arch.,  Bd.  58,  1873. 
Pasteur,  Bull,  de  1'Acad.  de  Med.,  T.  9,  1880.  Eecklinghausen  and  Lankowski, 
Virchow's  Arch.,  Bd.  60,  1873.  Waldeyer,  Arch,  f .  Gynakologie,  iii.,  1872. 

GONORRHOEA. 

Arning,  Viertelj.  f.  Dermatol.  u.  Syph.,  S.  371,  1883.  Aufrecht,  Patho- 
logische  Mittheilungen,  1881;  Centralbl.  f.  d.  Med.  Wiss.,  Nr.  16,  1883. 
Bockhart,  Sitzungsbericht  d.  Phys.  Med.  Ges.  zu  Wiirzburg,  1882 ;  Viertelj.  f. 
Dermatol.  und  Syph.,  1883.  Bokai,  Allgem.  Med.  Centralzeitung,  Nr.  74, 
1880.  Bokai  and  Finkelstein,  Prager  Med.  Chir.  Presse,  1880.  Biicker,. 
Ueber  Polyarthritis  Gonorrhoica.  Diss.,  1880.  Bumm,  Der  Mikroorganismus 
der  Gonorrhoischen  Schleimhauterkrankungen,  1885.  Campona,  Italia  Medica, 
1883.  Chameron,  Progres  Medical,  43,  1884.  Eschbaum,  Deut.  Med.  Woch., 
S.  187,  1883.  Frankel,  Deut.  Med.  Woch.,  Nr.  2,  1883  ;  S.  22,  1885.  Haab, 
Der  Mikrokokkus  der  Blennorrhcea  Neonator,  1881.  Hirschberg  and  Krause, 
Centralbl.  f.  Pract.  Augenheilk.,  1881.  Kammerer,  Centralbl.  f.  Chirurgie, 
Nr.  4,  1884.  Krause,  Die  Mikrokokken  der  Blenorrhoea  Neonator,  1882. 
Kroner,  Naturforschervers.  in  Magdeburg,  Arch.  f.  Gyn.,  xxv.,  S.  109,  1884. 
Leistikow,  Charite-Annalen,  7  Jahrg.,  S.  750,  1882.  Lundstrom,  Studier  ofver 
Gonokokkus  :  Diss.  Helsingfors,  1885.  Martin,  Kech.  sur  les  Inflamm.  Metast. 
.  a  la  Suite  de  la  Gonorrhe"e,  1882.  Neisser,  Centralbl.  f.  d.  Med.  Wiss., 
Nr.  28,  1879;  Deutsche  Med.  Woch.,  1882.  Newberry,  Maryland  Med.  Journ.,. 
1883.  Petrone,  Rivista  Clin.,  No.  2.  Eeter,  Centralbl.  f.  d.  Med.  Wiss.,  1879. 
Sanger,  Naturforschervers.  in  Magdeburg  ;  Ibid.,  S.  126,  1884.  Schrb'tter  and 
Winkler,  Centralbl.  f.  Bact,,  Bd.  ix.  Smirnoff,  Vrach.,  1886.  Steinschneider, 
Verhandl.  der  Deutsch,  Dermat.  Gesellsch.,  1889;  Berl.  Klin.  Woch.,  1890, 
Sternberg,  Med.  News,  Vol.  45,  Nr.  16,  1884.  Weiss,  Le  Microbe  du  Pus 
Blennorrhagique,  1889.  Welander,  Gaz.  Med.  de  Paris,  1884. 

OPHTHALMIC  DISEASES. 

Balogh,  Med.  Centralbl.,  xiv.,  1879.  Bock,  Virchow's  Arch.,  Bd.,  91,  1883. 
Cornil  and  Berlioz,  Compt.  Rend,  de  1'Acad.  d.  Sc.,  1883.  Deutschmann, 
v.  Graefe's  Archiv,  Bd.  xxxi.,  1885.  Gifford,  Archiv  f.  Augenheilkunde,  1886. 
Soldzieher,  Centralblatt  f .  Prakt.  Augenheilk.,  1884.  Kahler,  Prager  Zeitschr. 
f.  Prakt.  Heilk.,  Bd.  I.,  1882.  Klein,  Centralbl.  f.  d.  Med.  Wiss.,  Nr.  8,  1884. 


BIBLIOGRAPHY.  649 

Kroner,  Verb.  d.  Naturforscher-Vers.  Magdeburg,  1884.  Kuschbert,  Deutsch. 
Med.  Wocb.,  Nr.  21,  1884.  Kuschbert  and  Neisser,  Bresl.  Aerztl.  Zeitschr., 
Nr.  4,  1883.  Michel,  Graefe's  Arcbiv  f.  Augenbeilkunde,  1882.  Neisser, 
Fortschr.  d.  Med.,  Bd.  2,  S.  73,  1884.  Eeuss,  Wien.  Med.  Presse,  1884. 
Both,  Virchow's  Archiv,  Bd.  55,  1872.  Salomonsen,  Fortscbr.  d.  Med.,  Bd.  2, 
S.  78,  1884.  Sattler,  Ber.  lib.  d.  Ophthalmologen  Congress  zu  Heidelberg, 
1882;  Zehender's  Klin.  Monatsblatt,  and  Wien.  Med.  Wocb.,  Nr.  17,  1883. 
Sattler  and  de  Wecker,  L'Ophtbalmie  Je"quiritique,  1883.  Schleich,  Verb,  des 
Ophthalmologen  Congr.  zu  Heidelberg,  1883.  Vennemann  and  Bmylants,  Le 
Jequirity  et  son  Principe  Pathogene,  1884.  Vossius,  Berl.  Klin.  Wochenschr., 
Nr.  17,  1884.  Widmark,  Hygeia :  Stockholm,  1885.  Zehender,  Bowman 
Lecture  :  Lancet,  1886. 


CHAPTER    XIV. 

ANTHRAX. 

Archangelski,  Centralbl.  f.  d.  Med.  Wiss.,  1882,  1883.  Bert,  Compt.  Rend. 
Soc.  de  Biol.,  T.  4,  1877  ;  T.  5,  1878 ;  T.  6,  1879.  Bleuler,  Correspondenzbl.  d. 
8chweiz  Aerzte,  1884.  Bollinger,  Arbeit,  a.  d.  Patholog.  Inst.  zu  Miinchen, 
1886;  Centralbl.  f.  d.  Med.  Wiss.,  Bd.  10,  1872;  Sitzungsber.  d.  Ges.  f. 
Morphol.  Physiol.  zu  Miinchen,  1885.  Bouley,  Bull.  Acad.  de  Me"d.  :  Paris, 
T.  9,  1880  ;  T.  10, 1881 ;  Compt.  Eend.,  T.  92  and  93,  1881.  Brauell,  Virchow's 
Arch.,  Bd.  11,  1857;  Bd.  14,  1858.  Biichner,  Ueber  die  Exper.  Erzeugung 
des  Milzbrandcontagiums  aus  den  Heupilzen,  1880 ;  Sitzungsber.  d.  K.  Bayer. 
Akad.  d.  Wissenscb.,  1880;  Vortrage  im  Aerztl.  Verein  zu  Miinchen,  1881; 
Virchow's  Arch.,  Bd.  91,  1883.  Chauveau,  Compt.  Rend.,  T.  90  and  91,  1880; 
T.  92,  1881;  T.  94, 1882  ;  T.  96,  1883.  Chelchowsky,  Der  Thierarzt,  1884.  Colin, 
Bull.  Acad.  de  Med.  :  Paris,  T.  2,  1873 ;  T.  7,  1878 ;  T.  8,  1879  ;  T.  9,  1880 ; 
T.  10,  1881.  Crookshank,  Rep.  Agric.  Dept.,  1888.  Davaine,  Compt.  Rend. : 
Paris,  T.  77,  1873  ;  T.  57  and  59,  1863  ;  T.  60,  1865  ;  T.  61,  1866, 1877  ;  Rec.  de 
Med.  Vet.,  T.  4, 1877.  Dowdeswell,  Rep.  Med.  Off.  Local  Gov.  Board,  1883.  Esser 
and  Schiitz,  Mitth.  a  K.  Preuss  Amtl.  Vet.  Sanitatsbericht,  1882.  Ewart, 
Quart.  Journ.  of  Microsc.  Sc.,  1878.  Feltz,  Compt.  Rend.,  T.  95,  1882.  Fodor, 
Deut.  Med.  Woch.,  1886.  Fokker,  Centralbl.  f.  d.  Med.  Wissensch.,  Bd.  18, 
1880 ;  Centralbl.  f.  d.  Med.  Wiss.,  1881  ;  Virchow's  Archiv,  Bd.  88,  1882. 
Frank,  Zeitschr.  f.  Hygiene,  1886.  Friedrich,  Zur  Aetiologie  des  Milzbrands., 
1H85.  Greenfield,  Quart.  Journ.  Micr.  Sc.  London,  1879 ;  Proc.  Roy.  Soc. 
London,  1880.  Hoffa,  Die  Natur  des  Milzbrandgiftes :  Wiesbaden,  1886. 
Huber,  Deut,  Med.  Woch.,  Bd.  7,  1881.  Johne,  Ber.  ii.  d.  Veter.  Wesen.  i.  K. 
Sachsen,  188(5.  Kitt,  Sitzungsb.  d.  Ges.  f.  Morphol.  u.  Physiol.  zu  Miinchen, 
1885.  Klein,  Rep.  of  the  Medical  Officer  of  the  Local  Govt.  Board,  1881 ;  Quart. 
Journ.  Micr.  Sc.,  1883.  Koch,  Beitrage  zur  Biologic  der  Pflanzen,  Bd.  II., 
Heft  2,  1876  ;  Wrundinfectionskrankheiten,  1878  ;  Mitth.  aus  d.  Ges.  Amt, 
Bd.  I.,  1881  ;  Milzbrand  und  Rauschbrand  :  Stuttgart,  1886.  Martin,  Proc.  Roy. 
Soc.,  1890  ;  Rep.  Med.  Off.  Local  Govt.  Board,  1890-91 ;  Brit.  Med.  Journal, 
1891.  Oemler,  Archiv  f.  Wiss.  u.  Pract.  Thierheilk.,  Bd.  4,  1878,  1879,  1880. 
Osol,  Experiment.  Untersuch.  ii.  das  Anthraxgift:  Inaug.  Diss.  Dorpat,  1885. 
Pasteur,  Bull.  Acad.  de  M6d.,  1877,  1879, 1880  ;  Compt.  Rend.,  Paris,  T.  84, 1877  ; 
T.  90  and  91,  1880 ;  T.  92,  1881  ;  T.  95,  1882.  Pollender,  Vierteljahrschr. 
f.  Ger.  Med.,  Bd.  8,  1855.  Prazmowski,  Acad.  d.  Wissensch.  in  Krakau,  1884  ; 


•650  APPENDICES. 

Biol.  Centralbl.,  Bd.  4,  1884.  Eodet,  Contribution  &  1' Etude  Experimentelle 
du  Carbon  Bacteridien,  1881 ;  Compt.  Rend.,  T.  94,  1882.  EolofF,  Archiv  f. 
Wissensch.  u.  Bract.  Thierheilk.,  Bd.  9,  1883  ;  Der  Milzbrand  :  Berlin,  1883. 
Schmidt,  Deut.,  Zeitschr.  f.  Thiermed.  u.  Vergl.  Pathol.,  1879.  Schrakamp, 
Archiv  f.  Hygiene,  Bd.  2,  1884.  Semmer,  Centralbl.  f.  d.  Med.  Wissensch., 
Bd.  18,  1880,  1884  ;  Der  Milzbrand  und  das  Milzbrandcontagium,  1882. 
Steinberg,  Am.  Monthly  Micr.  Journ.,  1881.  Szpilman,  Zeitschr.  f.  Physiol. 
•Chemie:  Strassburg,  Bd.  4,  1880.  Toepper,  Die  Neueren  Erfahrungen  iiber 
d.  Aetiologie  d.  Milzbrands.,  1883.  Toussaint,  Compt.  Bend.,  T.  85,  1877, 
1878,  1880  ;  Eecherches  Experimentales  sur  la  Maladie  Charbonneuse,  1879. 
Wachenheim,  Etude  Experimentelle  sur  la  Septicite  et  la  Virulence  du  Sang 
Charbonneux,  1880. 


CHAPTER   XV. 

•QUARTER-EVIL.      MALIGNANT    (EDEMA.       RAG-PICKER'S    SEPTICAEMIA 
OF   GUINEA-PIGS.      SEPTIOEMIA   OF   MICE. 

QUARTER-EVIL. 

Arloing,  Cornevin  and  Thomas,  Compt.  Rend.,  1880 ;  Bull,  de  1'Acad.  de 
Med.,  and  Revue  de  Med.,  1881 ;  Du  Charbon  Bacterien,  Charbon  Symptoma- 
tique,  etc.,  1883 ;  Revue  de  Med.,  1884  ;  Chabert's  Disease  :  Transl.  by  Dawson 
Williams  in  Micro-parasites  and  Disease  (New  Syd.  Soc.),  1886.  Babes, 
Journ.  de  1' Anatomic,  1884.  Bellinger  and  Feser,  Wochenschr.  f.  Thierheil- 
kunde,  1878.  Ehlers,  Unters.  ub.  d.  Rauschbrandpilz :  Inaug.  Diss.  Rostock, 
1884.  Hess,  Bericht  iiber  die  entschadigten  Rauschbrand  u.  Milzbrandfalle 
im  Canton  Bern,  1886.  Kitt,  Jahresber.  der  K.  Thierarzneisch.  in  Miinchen, 
1884.  Neelsen  and  Ehlers,  Ber.  d.  Naturforsch.  Ges.  zu  Rostock,  1884. 
-Strebel,  Schweiz.  Archiv  f.  Thierheilk.,  1886. 

MALIGNANT  (EDEMA. 

Brieger  and  Ehrlich,  Berl.  Klin.  Wochenschr.,  N.  44,  1882.  Chauveau  and 
Arloing,  Archiv  Vet.,  1884 ;  Bull.  Acad.  de  Med.,  1884.  Davaine,  Bull,  de 
1'Acad.  de  Med.,  1862.  Gaffky,  Mitth.  as.  d.  K.  Ges.  Amt.,  1881.  Hesse,  W. 
and  R.,  Deut.  Med.  Woch.,  1885.  Kitasato  and  Weyl,  Zeitschr.  f.  Hygiene, 
Bd.  VIII.  Kitt,  Jahresber.  der  K.  Thierarzneischule  in  Miinchen,  1884.  Koch, 
Mitth.  aus  dem  Ges.  Amt,  I.,  S.  54,  1881.  Lebedeff,  Arch,  de  Phys.  Norm,  et 
Path.,  1882.  Lustig,  Jahresber.  d.  K.  Thierarzneischule  zu  Hannover,  1884. 
Pasteur,  Bull,  de  1'Acad.  de  Med.,  1877, 1881.  Roger,  Compt.  Rend.  Soc.  de  Biol., 
1889.  Roux  and  Chamberland,  Ann.  de  1'Institut  Pasteur,  1887.  Trifaud,  Rev. 
de  Chirurg,  T.  iii.  Van  Cott,  Centralb.  f.  Bact.,  1891.  Verneuil,  La  Semaine 
Med.,  1890. 

RAG-PICKER'S  SEPTIC^JMIA.    SEPTIC^MIA  OP  GUINEA-PIGS.    SEPTICAEMIA 

OF  MICE. 

Bordoni-Uffreduzzi,  Zeitsch.  f.  Hygiene,  1888.  Klein,  Centralbl.  f.  Bac- 
teriolog.,  1890.  Koch,  Wundinfectionskrankeit,  Trans.  New  Syd.  Soc.,  1880. 
Paltauf,  Wiener  Klin.  Woch.,  1888. 


BIBLIOGRAPHY.  651 

CHAPTER  XVI. 

ILEMORRHAGIC   SEPTICAEMIA. ' 

SEPTICAEMIA  OP  BUFFALOES.  SEPTIC  PLEUBO-PNEUMONIA  OF  CALVES. 
SWINE  FEVER.  SEPTICAEMIA  OF  DEEE.  SEPTICAEMIA  OF  RABBITS. 
FOWL  CHOLERA.  FOWL  ENTERITIS.  DUCK  CHOLERA.  GROUSE  DISEASE. 

Babes,  Compt.  Rend,  de  1'Acad.  d.  So.,  1883;  Arch,  de  Phypiol.,  1884. 
Barthelemy,  Compt.  Rend.,  T.  96,  No.  18,  1883.  Bunzl-Federn,  Centralbl.  f. 
Bacteriolog.,  1891.  Camera,  Centralbl.  f.  Bacteriolog.,  1891.  Cornil,  Arch,  de 
Physiol.,  Bd.  10,  1882.  Cornil  and  Chantemesse,  Le  Bulletin  Med.,  1887. 
Cornil  and  Toupet,  Bull,  de  la  Soc.  Nat.  d'Acclimation.,  1888.  Davaine, 
Bull,  de  1'Acad.  de  Med.,  1879.  Eberth  and  Schimmelbusch,  Virchow's 
Archiv,  1889  ;  Fortschr.  d.  Med.,  Bd.  VI.  Gaffky,  Mitt,  aus  dem  K.  G.  Amte, 
1881.  Gamaleia,  Centralb.  f.  Bacteriolog.,  1888.  Hueppe,  Berl.  Klin.  Woch., 
1886.  loannes  and  Megnin,  Journ.  d'Acclimatation,  1877.  Kitt,  Allg.  Deut. 
Geflugelzeitung,  1885.  Klein,  Rep.  Med.  Off.  Local  Govt.  Board,  1877-78; 
Fortschr.  der  Med.,  1888;  Centralb.  f.  Bakteriolog.,  1889.  Koch,  Aetiologie 
der  Wundinfections  R.,  1878.  Oreste  and  Armani,  Atti  de  R.  Instit.  d'Incorrag. 
Alle  Sci.  Nat.,  Econ  e  Tech.,  1887.  Pasteur,  Compt.  Rend.,  T.  90,  1886. 
Perroncito,  Arch.  f.  Wiss.  u.  Prakt.  Thierheilk.,  1879.  Petri,  Centralbl.  f.  d. 
Med.  Wiss.,  1885.  Bietsch  and  Jobert,  Compt.  Rend.,  1888.  Salmon,  Reports 
Bureau  of  Animal  Industry,  1886,  1887,  1888.  Salmon  and  Smith,  Amer. 
Monthly  Med.  Journ.,  1881.  Schiitz,  Archiv  f.  Wiss.  und  Prakt.  Thierheilk.,  1888. 
Semmer,  Deut.  Zeitschr.  f.  Theirmed.  u.  Vergl.  Path.,  1878.  Smith,  Journ.  f. 
-Comp.  Med.  and  Surg.,  1887 ;  Zeitschr.  f .  Hygiene,  1891.  Smith  and  Veranus 
Moore,  Rep.  Bureau  of  Animal  Industry,  1895.  Ziirn,  Die  Krankheiten  des 
Hausgeflugels,  1882. 


CHAPTER  XVII. 

PNEUMONIA.      INFECTIOUS   PLEURO-PNEUMONIA   OF   CATTLE. 
INFLUENZA. 

PNEUMONIA. 

Afanassiew,  Compt.  Rend.  Soc.  de  Biol.  Paris,  T.  5, 1884.  De  Blasi,  Rivista 
Internaz.  di.  Med.  e  Cbir.,  1885.-  Bruvlant  and  Verriers,  Bull,  de  1'Acad. 
Beige,  1880.  Dreschfeld,  Fortschr.  d.  Med.,  Bd.  3,  389,  1885.  Foa  and 
Bordoni-TJffreduzzi,  Deut.  Med.  Woch.,  1886.  Frankel,  Verhandl.  d.  Congr. 
f.  Innere  Med.,  Fortschr.  d.  Med.  Nov.,  1884;  Deut.  Med.  Woch.,  1886; 
Zeitschr.  f.  Klin.  Med.,  Bd.  x.  and  xi.,  1886.  Friedlander,  Virchow's  Arch., 
Bd.  87,  1882 ;  Fortschr.  d.  Med.,  Bd.  I.,  1883 ;  Bd.  II.,  1884 ;  Bd.  3,  92,  1885. 
Friedlander  and  Frobenius,  Berl.  Klin.  Woch.,  1883.  Germain-Se*e,  Compt. 
Rend.  Acad.  de  Sc.  Paris,  1884 ;  Des  Maladies  Specifiques  du  Poumon,  1885. 
<Jiles,  Brit.  Med.  J.,  Vol.  II.,  1883.  Griffini  and  Cambria,  Centralbl.  f.  d.  Med. 
Wiss.,  1883.  Jaccoud,  La  France  Medicale,  1886.  Jiirgensen,  Berl.  Klin. 
Woch.,  Bd.  22,  1884.  Klein,  Centralbl.  f.  d.  Med.  Wissensch.,  1884.  Koranyi 
and  Babes,  Pester  Med.  Chir.  Presse,  1884.  Ktthn,  Deutsch.  Arch.  f.  Klin. 
Med.,  1878  ;  Berl.  Klin.  Woch.,  Nr.  38,  1881.  Lancereaux  and  Besancon, 


OF  THK 

UNIVERSITY 


652  APPENDICES. 

Archiv  Gen.  de  M6d.,  1886.  Maguire,  Brit.  Med.  Journ.,  Vol.  II.,  1884. 
Manfredi,  Fortsch.  d.  Med.,  1886.  Matray,  Wien.  Med.  Presse,  Nr.  23,  1883. 
Mendelssohn,  Zeitschr.  f.  Klin.  Med.,  Bd.  7,  1884.  Nauwerck,  Beitr.  zur 
Pathol.  Anat.  von  Ziegler,  1884.  Neumann,  Berl.  Klin.  Woch.,  1885.  Paw- 
lowsky,  Berl.  Klin.  Woch.,  1885.  Peterlein,  Bericht  u.  d.  Vet.-Wesen.  i.  K. 
Sachsen,  1885.  Pipping,  Fortsch.  d.  Med.,  Nos.  10  and  14,  1886.  Platanow, 
Mitth.  a.  d.  Wiirzburg.  Med.  Klinik,  1885;  Ueber  die  Diagnostische 
Bedeutung  d.  Pneumoniekokken  :  Inaug.-Diss.  Wiirzburg,  1884.  Eibbert, 
Deut.  Med.  Woch.,  Nr.  9,  1885.  Salvioli  and  Zaslein,  Centralbl.  f.  d.  Med. 
Wissensch.,  1883.  Arch,  pour  les  Sc.  Med.,  T.  8.,  1884.  Schou,  Fortschr.  der  Med.r 
Bd.  3,  Nr.  15,  1886.  Sternberg,  Amer.  Journ.  Med.  Sciences,  1885;  Journ. 
Koy.  Micr.  Soc.,  1886.  Talamon,  Progr.  Medic.,  1883.  Thost,  Deut.  Med. 
Woch.,  1886.  Weichselbaum,  Wien.  Med.  Woch.,  1886.  Ziehl,  Centralbl.  f.  d. 
Med.  Wiss.,  1883;  Centralbl.  f.  d.  Med.  Wiss.,  1884. 

CEREBRO-SPINAL  MENINGITIS. 

Bonome,  Centralbl.  f.  Bact.  u.  Parasitolog.,  IV.  Foa,  Zeitschr.  f.  Hygiene, 
IV.  Leichtenstern,  Deut.  Med.  Woch.,  Nr.  23  u.  31,  1885.  Leyden,  Centralbl. 
f.  Klin.  Med.,  Nr.  10,  1883.  Weichselbaum,  Wien.  Klin.  Woch.,  1888. 

PLEURO-PNEUMONIA. 

Arloing,  Compt.  Kend.  cix.,  1889.  Bruce  and  Loir,  Ann.  de  1'Institut 
Pasteur,  1891,  Bruylants  and  Verriers,  Bull,  cle  1'Acacl.  Belg.,  1880.  Cornil 
and  Babes,  Arch,  de  Physiol.  Norm,  et  Path.,  T.  2,  1883.  Lustig,  Centralb.  f. 
die  Med.  Wiss.,  1885.  Mayrwieser,  Woch.  f.  Thierheilk.  u.  Viehzucht,  19,  1884. 
Pasteur,  Recueil  de  Med.  Vet.,  1882.  Poels,  Fortsch.  d.  Med.,  1886.  Poels 
and  Nolen,  Centralbl.  f.  d.  Med.  Wiss.,  Nr.  9,  1884 ;  Fortsch.  d.  Med.,  1886. 
Putz,  Thier.  Med.  Vortrage,  Bd.  1,  1889.  Eeport  on  Pleuro-pneumonia  and 
Tuberculosis,  1888.  Schutz  and  Steffen,  Archiv  f .  Wiss.  und  Prakt.  Thierheilk., 
Bd.  xv.  Sussdorff,  Deutsche  Zeitschr.  f.  Thiermed.  u.  Vergl.  Pathol.,  1879. 

INFLUENZA. 

Babes,  Centralbl.  f.  Bacteriolog.,  1890;  Deutsche  Med.  Woch.,  1892.  Bein, 
Zeitschr.  f.  Hygiene,  1890.  Bouchard,  La  Semaine  Med.,  1890.  Canon,  Deutsche 
Med.  Woch.,  1892.  Fischel,  Prager  Med.  Woch.,  1890 ;  Zeitschr.  f.  Heilkunde, 
1891.  Jolles,  Wiener  Med.  Blatt,,  1890.  Kirchner,  Centralbl.  f.  Bacteriolog., 
1890;  Zeitschr.  f.  Hygiene,  1890.  Kitasato,  Deutsche  Med.  Woch.,  1892.  Klein, 
Brit,  Med.  Journ.,  1892.  Klebs,  Centralbl.  f.  Bakteriolog.,  1890  ;  Deutsche  Med. 
Woch.,  1890.  Pfeiffer,  Deutsche  Med.  Woch.,  1892.  Prudden,  New  York  Med, 
Rec.,  1890.  ' 

CHAPTER    XVIII. 

ORIENTAL   PLAGUE.     RELAPSING   FEVER.      TYPHUS   FEVER.      YELLOW 

FEVER. 

ORIENTAL  PLAGUE. 

Aoyama,  Mitth.  u.  d.  Pest.  Epidemic  im  Jahre  1894;  in  Hong  Kong,  1895. 
Yeriin,  Ann.  de  1'Institut  Pasteur,  1894.  Yersin,  Calmette  and  Borrel,  Ann. 
de  1'Institut  Pasteur,  1895. 


BIBLIOGRAPHY.  653 

RELAPSING  FEVBE. 

Albrecht,  St.  Petersb.  Med.  Woch.,  1879.  Carter,  Lancet,  1879  ;  Trans. 
Internat.  Med.  Congress,  1881.  Cohn,  Deut.  Med.  Woch.,  1879.  Engel,  Berl. 
Klin.  Woch.,  1873.  Giinther,  Fortschr.  d.  Med.,  1885.  Guttmann,  Virchow's 
Arch.,  1880.  Heydenreich,  St.  Petersb.  Med.  Woch.,  1876  ;  Der  Parasit  des  Riick- 
falltyphus,  1877.  Jaksch,  Wien.  Med.  Woch.,  Juli,  1880.  Koch,  Deut.  Med. 
Woch.,  1879.  Laptschinsky,  Centralbl.  f.  d.  Med.  Wiss.,  Bd.  13,  1875.  Manas- 
sem,  St.  Petersb.  Med.  Woch.,  No.  18,  1876.  Metchnikoff,  Virchow's  Archiv, 
1877.  Moczutowsky,  Deutsches  Archiv  fiir  Klin.  Med.,  Bd.  xxiv.,  1876. 
Miihlhauser,  Virchow's  Arch.,  Bd.  97,  1880.  Obermeier,  Med.  Centralbl.,  11  ; 
Berl.  Med.  Ges. ;  Berl.  Klin.  Wochenschr.,  1873.  Soudakewitch,  Ann.  de 
I'lnstitut  Pasteur,  1891.  Weigert,  Deut.  Med.  Woch.,  1876. 

YELLOW  FEVEK. 

Babes,  "Compt.  Rend.,  17  Sept.,  1883.  Bouley,  Compt.  Rend.,  T.  100,  p.  1276, 
1885.  Carmona  y  Valle,  Lecons  sur  1'Etiol.  et  la  Prophylax.  de  la  Fievre 
Jaune,  1885.  Cerecedo,  El  Siglo  Medico,  1886.  Domingos  Freire,  Recherches 
sur  la  Cause  de  la  Fievre  Jaune,  1884 ;  La  Fievre  Jaune  et  ses  Inoculations 
Preventives,  1896.  Domingos  Freire  and  Rebourgeon,  Compt.  Rend.,  T.  99, 

p.  804,  1884. 

• 

CHAPTER    XIX. 

SCARLET   FEVER   AND   MEASLES. 

SCAKLET  FEVER. 

Coze  and  Feltz,  Les  Maladies  Infectieuses,  1872.  Crooke,  Lancet,  1883 ; 
Fortsch.  d.  Med.,  1885.  Crookshank,  Report  Agric.  Dept.,  1887.  Halm,  Berl. 
Klin.  Woch.,  No.  38,  1882.  Heubner  and  Bahrdt,  Berl.  Klin.  Woch.,  Nr.  44, 
1884.  Klein,  Nature,  xxxiv.,  1886;  Report  of  the  Medical  Officer  of  the 
Privy  Council,  1887,  1888,  1889.  Laure,  Lyon  Medical,  1886.  McKendrick, 
Brit.  Med.  Journal,  1872.  Pohl-Pincus,  Centralbl.  f.  d.  Med.  Wiss.,  No.  36, 
1883.  Both,  Munch.  Aerztl.  Intelligenzbl.,  1883. 

MEASLES. 
Cornil  and  Babes,  Archiv  de  Phys.,  1883.    Keating,  Phil.  Med.  Times,  1882. 


CHAPTER    XX. 

SMALL-POX.      CATTLE   PLAGUE. 
SMALL-POX. 

Chauveau,  Compt.  Rend.,  1868.  Cohn,  Virchow's  Archiv,  Bd.  55,  1872. 
Copeman,  Brit.  Med.  Journ.,  1896  ;  The  Practitioner,  1896.  Cornil  and  Babes, 
Soc.  Med.  des  Hop.,  1883.  Crookshank,  History  and  Path,  of  Vaccination,  1889. 
Haccius,  Variola-vaccine,  1892.  Hamerink,  Ueber  die  sog.  Vac.  u.  Variola,  1884. 
Ischamer,  Aerztl.  Verein.  Steiermark,  1880.  Klebs,  Arch.  f.  Exp.  Path.  u. 
Pharmakol.,  Bd.  10,  1874.  Klein,  Rep.  Med.  Off.  Loc.  Govt.  Board,  1893-4. 


654  APPENDICES. 

Luginbuhl,  Verhdl.  d,  Phys.  Med.  Ges.  in  Wiirzburg,  1873.  Marchand,  Revue 
Mycologique,  1882.  Pfeiffer,  Die  Protozoen  als  Krankheitserreger,  1890 ;. 
Behandl.  und  Prophylax.  der  Slattern,  1893.  Pissin,  Berl.  Klin.  Woch.,  1874. 
Plant,  Das  Organis.  Contagium  der  Schafpocken,  1883.  Pohl-Pincus,  Vaccina- 
tion, 1882.  Quist,  St.  Petersburg  Med.  Woch.,  Nr.  46,  1883.  Beports,  Royal 
Vaccination  Commission,  1888-96.  Buffer  and  Plimmer,  Brit.  Med.  Journ.,. 
1894.  Weigert,  Ueber  Bakterien  in  der  Pockenhaut,  1871 ;  Anat.  Beitr.  z. 
Lebre  v.  d.  Pocken,  1874.  Wolf,  Berl.  Klin.  Woch.,  Nr.  4.  1883.  Ziilzer,  Berl. 
Klin.  Woch.,  1872. 

CATTLE  PLAGUE. 

Crookshank,  History  and  Pathology  of  Vaccination,  1889.  Report  of  the 
Cattle  Plague  Commission,  1865.  Report  on  Indian  Cattle  Plague,  1871. 
Semmer  and  Archangelski,  Ueber  das  Rinderpestcontagium  und  dessen  Miti- 
gation ;  Centralbl.  f.  d.  Med.  Wiss.,  1883.  Simpson,  Brit.  Med.  Journal,  1896. 
Smart,  Reports  on  Cattle  Plague,  Edinburgh,  1865. 


CHAPTER    XXI. 

SHEEP-POX.      FOOT  AND   MOUTH   DISEASE. 

SHEEP-POX. 
Crookshank,  History  and  Path,  of  Vaccination,  1889. 

FOOT  AND  MOUTH  DISEASE. 

Klein,  Report  Med.  Off.  Local  Govt.  Board,  1885.     Schottelius,  Centralb.  f.- 
Bakteriolog.,  1892. 

CHAPTER    XXII. 

HORSE-POX.      COW-POX. 

HORSE-POX. 
Crookshank,  History  and  Pathology  of  Vaccination,  1889. 

COW-POX. 

Bucknill,  Prov.  Med.  Journ.,  1895.     Crookshank,  Brit.  Med.  Journ.,  1888; 

story  and  Pathology  of  Vaccination,  1889  (Vol.  II.  contains  reprints  of  the 

cs  of  Tenner,  Pearson,  Woodville,  Loy,  Rogers,  Birch,  Bousquet,  Estlin, 

Ceely,  Badcock,   Auzias-Turenne,  Dubreuilh,  Layet).     Eeports  of  the   Royal 

vaccination  Commission. 

CHAPTER    XXIII. 

DIPHTHERIA. 

«otbb0»  ?""'  J°hnS  H°Pkins  Univ"  1891,  1893  ;  Journ.  of  Path,  and  Back, 

on     -pi'   Virohow'8    Archiv-    1890.      Behring,    Deutsche   Med.    Woch. 

90.     Behring  and  Kitasato,  Deutsche  Med.  Woch.,  1890.     Birch-Hirschfeld, 

Archiv  fur  Heilk.,   1872.    Brieger   and  Frankel,    Berl.    Klin     Woch     1890. 


BIBLIOGRAPHY.  655- 

Buhl,  Zeitschr.  fur  Biol.,  1867.  Cornil,  Arch,  de  Physiol.,  1881.  Eberth,  Med.. 
Centralbl.,  XI.,  Nr.  8,  1873.  Emmerich,  Compt.  Rendus  et  Memoires  du  V. 
Congres  Internat.  d'Hygiene,  1884  ;  Deut.  Med.  Woch.,  1884.  Everett,  Med. 
and  Surg.  Reporter,  1881.  Forster,  Wien.  Med.  Woch.,  1881.  Francotte,  La 
Diphtheric,  1883.  Freidberger,  Deut.  Zeitschr.  fur  Thiermed.u.  Vergl.  Pathol.,. 
1879.  Fiirbringer,  Virch.  Arch.,  Bd.  91,  1883.  Gerhardt  and  Klebs,  Verhandl. 
d.  Congresses  1  Inn.  Med.,  1882,  1883.  Heubner,  Die  Experimentelle  Diph- 
theric, 1883.  Hueter  and  Tommasi,  Centralbl.  f.  d.  Med.  Wiss.,  1868.  Klebs, 
Arch.  f.  Exp.  Pathol.,  Bd.  4,  1875.  Klein,  Report  Med.  Dept.  Local  Govt. 
Board,  1889.  Letzerich,  Berl.  Klin.  Woch.,  xi.,  1874  ;  Virch.  Arch.,  Bd.  55,. 
1872 ;  Bd.  68,  1876.  Loffler,  Mittheil.  a.  d.  Kais.  Ges.  Amt,  Bd.  II.,  1884  ; 
Microparasites  and  Disease  (New  Syd.  Soc.),  1886  ;  Centralbl.  f.  Bacteriolog., 
1887,  1890;  Deutsche  Med.  Woch.,  1890.  Lumner,  Aerztl.  Int.  BL,  No.  31, 
1881.  Martin,  Rep.  Med.  Off.  LOG.  Gov.  Board,  1890.  Nenmayer,  Neue 
Thesen  zu  Diphtheritisfrage,  1880.  Nicati,  Compt.  Rend.,  T.  88,  1879.  Oertel, 
Deut.  Arch.  f.  Klin.  Med.,  Bd.  8,  1871 ;  Zur  Aetiologie  der  Infectionskrank- 
heiten,  1881.  Park,  New  York  Med.  Record,  1892,  1893.  Prudden,  Amer. 
Journal  of  Med.  Sci.,  1889  ;  New  York  Med.  Rec.,  1891.  Kivolta,  Giornale  de 
Anat.  Fisiol.  e  Patol.  delli  Anim.,  1884.  Boux  and  Yersin,  Ann,  de  1'Institut 
Pasteur,  1888,  1889,  1890.  Salisbury,  Gaillard's  Med.  Journ. :  New  York,  1882. 
Talamon,  Bull,  de  la  Soc.  Anat.  de  Paris,  T.  56,  1881.  Welch,  Amer.  Journal 
of  the  Med.  Sci.,  1894.  Welch  and  Abbott,  Bull.  Johns  Hopkins  Univ.,  1891. 
Wood  and  Formad,  Bull.  Nat.  Board  of  Health,  Wash,  and  Med.  Times  and 
Gazette,  1882.  Zahn,  Beitrage  zur  Pathol.  u.  Histol.  der  Diphtheric,  1878. 
Zarniko,  Centralbl.  f.  Back,  1889. 


CHAPTER    XXIV. 

TYPHOID   FEVER. 

Almquist,  Typho'idfeberus-Bakterie,  1882.  Beumer  and  Peiper,  Zeitschr.  f. 
Hygiene,  1886.  Birch-Hirschfeld,  Zeitschr.  f.  Epidemiologie,  I.,  1874.  Boens,. 
Acad.  Roy.  de  Med.  de  Belgique :  Bull.  3  Ser.,  T.  17,  1883.  Brautlecht, 
Virchow's  Arch.,  Bd.  84,  1881.  Coats,  Brit.  Med.  Journ.,  1882.  Crooke,  Brit. 
Med.  Journ.,  1882.  Eberth,  Arch.  f.  Pathol.  Anat.,  Bd.  81,  1880;  Volkmann, 
Klin.  Vortrage,  1883.  Eppinger,  Beitr.  zur  Pathol.  Anatomic  aus  d.  Patholog, 
Institut.  Prag.,  1880.  Feltz,  Compt.  Rend.,  T.  85,  1877.  Fischel,  Beitr.  zur 
Pathol.  Anat.  aus  d.  Pathol.-Anat.  Inst.  Prag.,  1880.  Fraenkel  and  Simmonds, 
Die  Aetiologische  Bedeutung  des  Typhus-Bacillus,  1886.  Gaff  ky,  Mitth.  a.  d.. 
Ges.  Amt,  Bd.  II.,  1884.  Klebs,  Arch.  f.  Exper.  Pathol.  u.  Pharmakol.,  1880. 
Klein,  Med.  Centralbl.,  xii.,  1874.  Letzerich,  Virchow's  Archiv,  Bd.  68, 1876  ,-. 
Archiv  f.  Exper.  Pathol.,  Bd.  9,  1878  ;  Bd.  10,  1881 ;  Experimentelle  Unter- 
suchungen  liber  die  Aetiologie  des  Typhus  Abdominalis  :  Leipzig,  1883.  Luca- 
tello,  Bollet.  d.  R.  Accad.  Med.  di  Geneva,  1886.  Maraghano,  Centralbl.  f.  d. 
Med.  Wiss.,  Bd.  15,  1882.  Meisels,  Wien.  Med.  Woch.,  1886.  Meyer,  Unters. 
iiber  den  Bacillus  des  Abdominaltyphus  :  Inaug.  Diss.,  1881 .  Michael,  Fortsch. 
d.  Med.,  1885.  Neuhauss,  Berl.  Klin.  Woch.,  1886.  Pfeiffer,  Deut.  Med. 
Woch.,  Nr.  29,  1885.  Eappin,  Contrib.  a  1' Etude  des  Bact.  de  la  Bouche, 
a  1'Etat  Normal  et  dans  la  Fievre  Typho'ide,  1881.  Seitz,  Bakteriolog.  Studiea 
z.  Typhusatiologie,  1886.  Sirotinin,  Zeitsch.  f.  Hygiene,  1886.  Tayon,  Compt. 
Rend.,  T.  99,  p.  331,  1884.  Tizzoni,  Studi.  di  Pat.  Sperim.  sulla  Gen.  d.  Tifo.r- 
Milan,  1880.  Wernich,  Zeitschr.  f.  Klin.  Med.,  Bd.  6,  1882. 


OF  THK 

UNIVERSITY 


656  APPENDICES. 

CHAPTER   XXV. 

SWINE-TYPHOID. 

Klein,  Rep.  of  the  Med.  Offic.  of  the  Privy  Council,  1877-78  ;  Virchow's 
Archiv,  1884.  M'Fadyean,  Journ.  of  Comp.  Path,  and  Therapeutics,  1895. 
Report  of  a  Conference  on  Swine-fever :  Board  of  Agriculture,  1896.  Rietsch, 
Jobert  and  Martinaud,  Compt.  Kend.,  T.  106.  Selander,  Centr.  f.  Bakt.  u. 
Parasitenk.,  1888  ;  Ann  de  1'Institut  Pasteur,  1890.  Smith  and  Veranus  Moore, 
Bull,  of  Bureau  of  Animal  Industry  U.S.,  1894.  Welch  and  Clement,  Trans. 
Internat.  Vet.  Congress  of  Amer.,  1893. 

CHAPTER   XXVI. 

SWINE-MEASLES.      DISTEMPER   IN   DOGS.      EPIDEMIC   DISEASE   OF 

FERRETS.      EPIDEMIC   DISEASE    OF   MICE. 
• 

SWINE-MEASLES. 

Cornil  and  Babes,  Arch,  de  Physiol.,  1883.  Detmers,  Science,  1881. 
Eggeling,  Fortschr.  d.  Med.,  1883.  Loftier,  Arbeiten  aus  dem  Kaiserl.  Gesund- 
heits  Amt,  Bd.  I.,  1885.  Lydtin  and  Schottelius,  Der  Rothlauf  der  Schweine  : 
Wiesbaden,  1885.  Pasteur,  Compt.  Rend.,  T.  95,  1882.  Pasteur  and  Thuillier, 
Bull,  de  1'Acad.  de  Med.  de  Paris  :  Compt.  Rend.,  T.  97,  1883.  Salmon, 
Report  Depart.  Agricul.  Washington,  1881,  1884.  Schiitz,  Ueber  den  Rothlauf 
der  Schweine  und  die  Impfung  desselben,  1885. 


CHAPTER   XXVII. 

ASIATIC  CHOLERA.  CHOLERA  NOSTRAS.  CHOLERAIC  DIARRHOEA 
FROM  MEAT-POISONING.  DYSENTERY.  CHOLERAIC  DIARRHOEA 
OF  FOWLS. 

CHOLEEA. 

Ali-Cohen,  Fortschr.  der  Med.,  1887,  1888.  Almquist,  Zeitschr.  f.  Hygiene, 
1887.  Babes,  Virchow's  Archiv,  1885.  Bianchi,  Lancet.,  1884.  Bitter,  Archiv  f. 
Hygiene,  1886.  Bochefontaine,  Exper.  pour  servir  a  1'Etude  des  Phenomenes 
determines  chez  1'Homme  par  1'Ingestion  Stomacale  du  Liquide  Diarrh6'ique 
du  Cholera  :  Compt.  Rend.,  1884.  Brieger,  Deutsche  Med.  Woch.,  1887 ;  Berl. 
Klin.  Woch.,  1887 ;  Virchow's  Archiv,  1887.  Brunetti,  Fatti  Considerazioni 
Conclusioni  sul  Colera,  1885.  Biichner,  Archiv  f.  Hygiene,  1885;  Miinchn.  Aerztl. 
Intelligenzbl.,  S.  549,  1884.  Biichner  and  Emmerich,  Munch.  Med.  Woch.,  1885. 
Bujwid,  Zeitschr.  f.  Hygiene,  Centralbl.  f.  Bacteriolog.,  1,888.  Cameron,  Brit. 
Med.  Journ.,  1884.  Cattani,  Deut.  Med.  Woch.  1886.  Carter,  Lancet,  1884. 
Cheyne,  Brit.  Med.  Journ.,  1885.  Crookshank,  Lancet,  1885.  Cunningham, 
Scientific  Memoirs  of  Med.  Off.  of  Indian  Army,  1885,  1891.  Deneke,  Deut. 
Med.  Woch.,  Nr.  3,  1885.  Doyen,  Soc.  de  Biol.  de  Paris,  1884.  Drasche, 
Allg.  Wien.  Med.  Zeit.,  1885.  Dunham,  Zeitsch.  f.  Hygiene,  1887.  Emmerich, 
Deut.  Med.  Woch.,  Nr.  50,  1884.  Ermengem,  Deut.  Med.  Woch.,  Nr.  29,  1885 ; 


BIBLIOGRAPHY.  657 

Kecherches  sur  le  Microbe  flu  Cholera  Asiatique,  1885.  Ferran,  Compt.  Rend., 
T.  KX),  p.  959,  1NS">.  Finkler  and  Prior,  Naturforscherversammlung  Magde- 
burg, 1884;  Deut.  Med.  Woch.,  Nr.  36,  1884;  Erganzungshefte  zum  Centralbl. 
f.  Allg.  Gesundheitspflege,  Bd.  I.,  Heft  5  u.  6,  1885.  Fliigge,  Deut.  Med. 
Woch.,  Nr.  2,  1885.  Gaffky,  Arb.,  aus  d.  K.  Gesundheitsamte,  1887.  Gamaleia, 
Compt.  Rend.,  1888;  Compt.  Rend.  Soc.  de  Biolog.,  1889.  Gibier  and  van 
Ermengem,  Compt.  Rend.,  T.  101,  1885.  Haffkine,  Brit.  Med.  Journ.,  1895. 
Hericourt,  Compt.  Rend.,  1885.  Hueppe,  Berl.  Klin.  Woch.,  1887  ;  Deutsche 
Med.  Woch.,  1887;  Compt.  Rend.,  1889;  Prag.  Med.  Woch.,  1889.  Hunter, 
Brit.  Med.  Journ.,  1884.  Johne,  Deutsche  Zeitschr.  f.  Thiermed.,  Bd.  XI.,  1885. 
Kitasato,  Zeitschrif,t  f.  Hygiene,  1888,  1889.  Klebs,  Ueber  Cholera  Asiatica, 

1885.  Klein,  Brit.  Med.  Journ.  and  Proc.  Roy.    Soc.   London,  No.  38,  1885  ; 
Bacteria  of  Asiatic  Cholera,  1889.     Klein  and  Gibbes,   An  Inquiry   into  the 
Etiology  of  Asiatic  Cholera  :  Bluebook,  1885.     Koch,  Deut.  Med.  Woch.,  1884  ; 
Etiology  of   Cholera :    Berlin  Cholera  Conference  :   Translated   by  Laycock, 
in  Microparasites  and  Disease  (New  Syd.  Soc.),    1885.      Lewis,    Med.  Times 
and  Gazette,  1884.      Lustig,  Centralbl.  f.  die  Med.  Wiss.,  1887  ;  Zeitsshrift  f. 
Hygiene,  1887.     Macnamara,   Brit.   Med.  Journ.,  1884.      Miller,  Deut.  Med. 
Woch.,  Nr.  9,  1885.     Neuhaus,   Centralbl.  f.  Bacteriolog.,  1889.     Nicati  and 
Rietsch,   Arch,   de   Physiol.,   1885:    Revue   de   Medecin,   T.  5,    1885;    Revue 
d'Hygiene,  1885.     Petri,  Centralbl.  f.  Bacteriolog.,  1889.     Pettenkofer,  Lancet, 

1886.  Pfeiffer,  Corresp.-Bl.  des  Allgem.  Aerztl.  Ver.  inThiiringen,  Nr.  9,  1884  ; 
Deut.  Med.  Woch.,  1886-8S.     Pfuhl,   Zeitschr.  f.  Hygiene,  1889.    Boy,  Brown 
jtad  Sherrington,  Proc.  Roy.  Soc.,  Vol.  xli.     Salkowski,  Virchow's  Arc'aiv,  1887. 
Schottelius,  Deut,  Med.  Woch.,  Nr.  14,  18S5.     Strauss,  EDUX,  Thuillier  and 
Nocard,    Compt.    Rend.    Soc.   de    Biol.,   T.   4,    1883.      Tizzoni    and    Cattani, 
Centralb.  f.  die  Med.  Wiss.,  1886,  1887.    Vincenzi,  Deutsche  Med.  Woch.,  1887. 
Wassiljew,  Zeitschr.  f.  Hygiene,  1887.     Weisser,  Zeits3hrift  f.  Hygiene,  1886. 
Weisser    and   Frank,    Zeitschr.   f.   Hygiene,    18S(5.     Z'islein,    Deutsche   Med. 
Zeitung,  1887-88. 


CHAPTER    XXVIII. 

TUBERCULOSIS. 

Albrecht,  Arch.  f.  Kinderheilk.,  Bd.  5,  1884.  Andrew,  Lancet,  1884. 
Arloing,  Compt.  Rend.,  1884.  Aufrecht,  Centralbl.  fur  d.  Med.  Wiss.,  1882.  1883. 
Babes,  Compt.  Rendus,  1883  ;  Centralbl.  f.  d.  Mel.  Wissensch.,  lss:{.  Babes 
and  Cornil,  Journ.  de  1'Anat.  et  de  la  Physiol.  Norm,  et  Pathol.,  1884.  Balogh, 
Wien.  Med.  Woch.,  1882.  Baumgarten,  Berl.  Klin.  Wo;h.,  Bd.  17.  1*79 ; 
Centralbl.  f.  d.  Med.  Wiss.,  Bd.  19.  1881  ;  Bd.  20,  1882  ;  Bd.  21,  1883  ;  Bd.  22, 
1884;  Deut.  Med.  Woch.,  Bd.  8,  1882;  Zeitschr.  f.  Klin.  Med.,  Bd.  6,  1883, 
1886.  Biedert,  Virchow's  Archiv,  Bd.  98,  1884  ;  Berl.  Klin.  Woch.,  18S.->.  Black, 
Lancet,  1886.  Bock,  Virchow's  Archiv,  Bd.  91,  1883.  Bellinger,  Centralbl.  f. 
d.  Med.  Wiss.,  Bd.  21,  S.  600,  1883  ;  Miinch.  Aerztl.  :  Intelligenzbl.,  Nr.  K-, 
iss:;.  Bouley,  La  Nature  Vivante  de  la  Contagion,  Contagiosite  de  la  Tuber- 
culose:  Paris,  1884.  ,  Brouilly,  Rev.  de  Chir.,  T.  3.,  1883.  Celli  and  Guarneri, 
Arch,  pour  les  Sciences  Medic.,  188:5.  Cheyne,  Brit.  Mi-d.  .lourn..  Vol.  I..  ;  -v;;  . 
Practitioner,  Vol.  XXX.,  1883.  Chiari,  Wirn.  .Mod.  Presse,  1883.  Cochet, 
Compt.  Rend.  Soc.  de  Biol.  :  Paris,  T.  5,  1883.  Cohnheim,  Uebertragbarkeit 
der  Tuberculose  :  Berlin,  1877.  Cornil  and  Leloir,  Arch,  de  Physiol.  Norm,  et 

42 


(J58  APPENDICES. 

Pathol.,  1884.  Cramer,  Sitzungsber.  d.  Phys.  Med.  Soc.  zu  Erlangcn,  1883. 
Creighton,  Trans.  Path.  Soc.,  1882  ;  Brit:  Med.  Journ.,  1885  ;  Lancet,  1885. 
Crookshank,  Rep.  Agric.  Dept,,  1888  ;  Proc.  Phys.  Soc.,  1890  ;  Trans.  Path.  Soc., 
1891.  Damsch,  Deut.  Med.  Woch.,  Nr.  17,  1883.  Dejerine,  Eev.  de  Med. :  Paris, 
T.  4  1884.  Demme,  Jahresber.  d.  Jennerschen  Kinderspitals  :  Bern,  1883.  Dett- 
weiler,  Berl.  Klin.  Woch.,  Bd.  21,  1880.  Dieulafoy  and  Krishaber,  Arch,  de 
Physiol.  Norm.  etPath.,  T.  I.,  1883.  Doutrelepont,  Vierteljahrschr.  f.  Dermato- 
logie  u.  Syphilis,  1884  ;  Deut.  Med.  Woch.,  Nr.  7,  1885.  Ehrlich,  Deut.  Med. 
Woch.,  1882,  1883.  Ermengem,  Ann.  de  la  Soc.  Beige  de  Microscopic,  1882. 
Ewart,  Lancet,  1882.  Formad,  The  Bacillus  Tuberculosis:  The  Philad. 
Medical  Times,  1882.  Frantzel  and  Palmers,  Berl.  Klin.  Woch.,  1882,  1883. 
Fiitterer,  Virch.  Arch.,  Bd.  100,  Heft  2,  1885.  Gaffky,  Mitth.  a.  d.  Kaiserl. 
Gesundheitsamte,  Bd.  II.,  1884.  Giacomi,  Fortschr.  d.  Med.,  Bd.  I.,  S.  145, 
1883.  Gibbes,  Lancet,  1883.  Goldenblum,  Vrach.,  Nos.  I.  and  XL,  1886. 
Green,  Brit.  Med  Journ.,  1883  ;  Lancet,  1887.  Harries  and  Campbell,  Lupus: 
London,  1886.  Harris,  St.  Barthol.  Hosp.  Reports,  1885.  Heron,  Lancet, 

1883.  Hiller,   Deut.   Med.  Woch.,  Bd.   8,  1882.     Johne,  Die  Geschichte  der 
Tuberculose,  1883  ;   Ber.   iib.  d.  Veterinarwesen  im  Kb'nigr  :   Sachsen,  1883  ; 
Fortschr.  d.  Med.,  Bd.  3,  198,  1885.     Karg,  Centralbl.  f.  Chir.,  1885.     Kirstein, 
Deut.  Med.  Woch.,  1886.     Klebs,  Virchow's  Arch.,  Bd.  44,  1868  ;  Arch.  f.  Exp. 
Pathol.  u.  Pharmakol.,  Bd.  I.,  1873  ;  Bd.  17,  1883.     Koch,  Die  Aetiologie  der 
Tuberculose:    Berl.   Klin.  Woch.,   1882;   Deut.   Med.   Woch.,  Nr.    10,   1883; 
Mittheilungen  aus  dem  Kais.  Ges.  Amt,  Bd.  II.,  1884.     Kundrat,  Wien.  Med. 
Presse,   1883.     Ktissner,   Deut.   Med.   Woch.,   Nr.   36,    1883.     Landouzy   and 
Martin,  Rev.  de  Med.,  T.  3,  1883.     Leube,  Sitzungsber.  der  Phys.-Med.  Soc.  zu 
Erlangen,  1883.    Levinsky,  Deut,  Med.  Woch.,  Bd.  9.,  1883.     Leyden,  Zeitschr. 
f.  Klin.  Med.,  VIII.,  1884.     Lichtheim,  Fortschr.  d.  Med.,  Bd.  I.,  1883.     Lustig, 
Wien.  Med.  Woch.,  Nr.  48,  1884.     Lydtin,  Badische  Thierarztl.  Mittheil,  1883. 
Malassez  and  Vignal,  Compt.  Rend.,  T.  97,  1883  ;   Cornpt.  Rend.,  T.  99,  p.  200, 

1884.  Marchand,  Deut.  Med.  Woch.,  Nr.  15,  1883.     Max-Bender,  Deut.  Med. 
Woch.,  1886.     Meisels,  Wien.   Med.  Woch.,  1883.     Middeldorpf,  Fortsch.  d. 
Med.,   1886.     Mtiller,    Centralbl.   f.    Chir.,  3,    1884,   1886.      Nauwerck,    Deut, 
Med.   Woch.,   1883.      Obrzut,   Deut.    Med.    Woch.,   Nr.    12.   1885.      Pfeiffer, 
Berlin.    Klin.    Woch.,   Bd.    21,    1883.      Piitz,    Ueber    die    Beziehungen   der 
Tuberculose  des  Menschen  zu  der  Thiere,   1883.     Kaymond,  Arch.  Gen.   de 
Med.,  T.  11,  1883.     Ribbert,  Deut.  Med.  Woch.,  1883,  1885.     Rindfieisch,  Phys. 
Med.  Ges.  zu  Wurzburg,  Nr.  8,  1882.    Eosenstein,  Centralbl.  f.  d.  Med.  Wiss., 
1883.     Schill  and  Fischer,    Mitth.  a.   d.   Kaiserl.   Ges.  Amt,   Bd.  II.,   1884. 
Schlegtendal,  Fortschr.  d.  Med.,  Bd.  I.,  1883.     Schottelius,  Virchow's  Archiv. 
Bd.  91,  1883.    Schuchardt  and  Krause,  Fortschr.  d.  Med.  1883.     Smith,  Bristol 
Med.  Chir.  Journ.,  1883.     Somari  and  Brugnatelli,  Redii  R.  Instit.  Lombardo, 
1883.     Spina,  Casopis  Lekaru  Ceskych,  Nr.  4,  1885  ;  Studien  iiber  Tuberculose  : 
Wien.,  1883.    Sticker,  Centralbl.  f.  Klin.  Med.,  1885.     Strassmann,  Virchow's 
Archiv,  Bd.  96,  1884.     Button,  Trans.  Path.  Soc.  London,  Vol.  XXXV.,  1884. 
Toussaint,  Compt.  Rend.,  T.93,  1881.     Tscherning,  Fortschr.  d.  Med.,  Bd.  3,  65, 

1885.  Veraguth,  Arch.  f.  Exp.  Path.  u.  Pharmakol.,  Bd.  16,  1883.    Vignal, 
Compt.  Rend.  Soc.  de  Biol.,  T.  5,  1883.    Villemin,  Etude  sur  la  Tuberculose, 

168.    Voltolini,  Deut.  Med.  Woch.,  Nr.  31.     Wahl,  Deut.  Med.  Woch.,  Nr.  46, 

Weichselbaum,  Wien.  Med.  Jahrb.,  1883.     Weigert,  Deut.  Med.  Woch., 

Nr.  24ff.  1883.    Wesener,  Fiitterungstuberculose,  1885.     West,  Lancet,  Vol.  I., 

I88o ;  Trans.  Path.   Soc.,  1883.     Williams,  Lancet.  1883  ;   Journ.   Roy.  Micr. 

Soc.,  1884.     Ziehl,  Deut.  Med.  Woch.,  Nr.  5.  1883. 


BIBLIOGRAPHY.  659 

CHAPTER    XXIX. 

LEPROSY.      SYPHILIS.      RHINOSCLEROMA.      TRACHOMA. 

LEPROSY. 

Arning,  Virchow's  Archiv,  Bd.  97,  1884.  Babes,  Compt.  Rend.,  1883.  Babes 
and  Kalindero,  La  Semaine  Med.,  1891.  Baumgarten,  Centralbl.  f.  Bact.. 
1887  ;  Berl.  Klin.  Woch.,  1889.  Beaven-Eake,  Trans.  Path.  Soc.,  1887  ; 
Reports  Leper  Asylum,  Trinidad.  Bordoni-Uffreduzzi,  Zeitschr.  f.  Hygiene 
1887  ;  Berl.  Klin.  Woch.,  1888.  Campava,  La  Reforma  Med.,  1889*  1891. 
Cornil  and  Suchard,  Ann.  de  Dermat.  et  Syph..  1881.  Creighton,  History 
of  Epidemics  in  Great  Britain,  1894.  Damsch,  Virch.  Arch..  Bd.  92 : 
Centralbl.  f.  d.  Med.  Wissensch.,  Bd.  21,  1883.  Gaucher  and  Hillairet, 
Progres  Med.,  1880.  Guttmann,  Berl.  Klin.  Woch.,  1885.  Hansen,  Virchow's 
Archiv,  1880,  1882,  1886.  Hillis,  Trans.  Path.  Soc.,  1883.  Hills,  On  Leprosy 
in  British  Guiana,  1881.  Kaposi,  Wiener  Med.  Woch.,  1883.  Kobner, 
Virchow's  Arch.,  1882  ;  Leloir,  Ann.  de  Dermat.  et  de  Syph.,  1887.  Lubimoff, 
Centralbl.  f.  Bacteriolog.,  1888.  Melcher  and  Ortmann,  Berl.  Klin.  Woch. 

1885,  1886.    Moretti,  II  Primo  Caso  di  Lebbra  nelle  Marcho  Conferraato  dalla 
Presenza  del  Bacillus  Leprae,  1883.     Miiller,  Deut.  Archiv  f.  Klin.  Med.,  Bd. 
34,  1883.     Neisser,  Breslauer  Aerztl.    Zeitschr.,  1879  ;   Jahresber.   d.  Schles 
Ges.  fur  Vaterl.  Cultur.,  1879  :  Virchow's  Archiv,  Bd.  84,  1881,  1886.    Newman, 
Leprosy  as  an  Endemic  Disease  in  the  British  Islands.  1895.     Report,  Leprosy 
Commission,    18<)3.      Steven,    Brit.   Med.    Journ.,    1885.      Thin,    Brit.     Med. 
Journ.,    1884.      Teuton,   Fortsch.  d.  Med.,  1886.     Unna,   Deut.  Med.  Woch.. 
Nr.  32, 1885,  1886  ;  Virchow's  Archiv,  1886.    TTnna  and  Lutz,  Dermatologische 
Studien,  Heft  I.,  1886.     Vidal,  La  Lepre  et  son  Traitement,  1884.     Virchow, 
Berl.  Klin.  Woch.,  N.  12,  1885.     Vossius,  Ber.  iiber  d.  Ophthalmologen  Con- 
gress in  Heidelberg,  1881.     Wesener,  Central,  f.  Bacteriolog..  1887  ;  Munch. 
Med.  Woch.,  1887. 

SYPHILIS. 

Alvarez  and  Tavel,  Bull,  de  1' Acad.  de  Med.  et  Archiv  de '  Phys.  Norm,  et 
Path.,  1885.  Aufrecht,  Centralbl.  f.  d.  Med.  Wissensch.,  Bd.  19,  1881. 
Bienstock,  Fortsch.  d.  Med.,  1886.  Birch-Hirschfeld,  Centralbl.  f.  d.  Med. 
Wissensch.,  Nrs.  33,  34,  1882.  Disse  and  Taguchi,  Deut.  Med.  Woch.,  1886. 
Doutrelepont  and  Schiitz,  Deut.  Med.  Woch.,  Nr.  19,  1885.  Eve  and  Lingard, 
Brit.  Med.  Journ.,  1886.  De  Giacomi,  Correspondenzbl.  f.  Schweizer  Aerzte, 
Bd.  15,  1885.  Gottstein,  Fortschr.  d.  Med.,  Bd.  3,  S.  543,  1885.  Heyden, 
Preservation  de  la  Syphilis,  etc.  :  Traduit  par  Roberts,  1883.  Kassowitz 
and  Hochsinger,  Wien.  Med.  Blatter,  1886.  Klebs,  Arch.  f.  Exp.  Pathol., 
Bd.  10,  1879.  Klemperer,  Deut.  Med.  Woch.,  1885.  Kbniger,  Deut.  Med. 
Woch.,  S.  816,  1884.  Letnik,  Wien.  Med.  Wochenschr.,  1883.  Lostorfer,  Arch, 
f.  Dermat.  u.  Syph.,  1872.  Lustgarten,  Wien.  Med.  Woch.,  Nr.  47,  1884  ;  Die 
Syphilisbacillen,  1885.  Martineau  and  Hamonio,  Compt.  Rend.,  p.  443,  1882. 
Morison,  Maryland  Med.  Journ.,  1882  ;  Ibid. :  Baltimore,  1883  ;  Wiener  Med. 
Wochenschr.,  18x3.  Peschel,  Centralbl.  f.  Augenheilk.,  1882.  Petrone,  Gaz. 
Medica  Ital.,  1884.  Torney  and  Marcus,  Compt.  Rend.,  p.  472;  1W4. 

RHINOSCLEROMA. 

Cornil,  B.  de  la  Soc.  Anatom.,  15  Fev.,  18S5.  Cornil  and  Alvarez,  Acad.  de 
Med.  et  Archiv  de  Phys.  Norm,  et  Path.,  18«5.  Davies,  Brit.  Med.  Journ  , 

1886.  Paltauf  and  Eiselsberg,  Fortsch.  d.  Med.,  1886. 


660  APPENDICES. 


CHAPTER    XXX. 

ACTINOMYCOSIS   AND   MADURA    DISEASE. 
ACTINOMYCOSIS. 

Acland,  Brit.  Med.  Journ.  and  Trans.  Path.  Soc.,  1886  ;  and  Allbut's  System  of 
Med.,  1896.  Bang,  Tidskrift  far  Veterinaerer,  1883.  Baumgarten,  Berl.  Klin. 
Woch.,  1885.  Bellinger,  Centralbl.  f.  d.  Med.  Wiss.,  1877.  Bostrdm,  Verb.  d. 
Congr.  f.  Inn.  Med.  Wiesbaden,  1888.  Chiari,  Prager  Med.  Woch.,  Nr.  10,  1884. 
Crookshank,  Report  of  the  Agric.  Dept.  of  the  Privy  Council,  1888  ;  Trans. 
Roy.  Med.  and  Chirurg.  Soc.,  1889.  Firket,  Rev.  de  Med.,  1884.  Fleming, 
Actinomycosis,  1883.  Gannet,  Boston  Med.  and  Surg.  Journ.,  1882.  Hertwig, 
Archiv  f.'  Wiss.  u.  Prakt.  Thierheilk;,  1886.  Hink,  Centralbl.  f.  d.  Med.  Wiss., 
1  ss2.  Israel,  Archiv  f.  Klin.  Chir.,  1868  ;  Virchow's  Arch.,  Bd.  74.  1878  ;  Bd.  78, 
1879  ;  Bd.  96.  1884  ;  Kenntniss  der  Actinomykose  des  Menschen  :  Klinische 
Beitrage,  1885.  Johne,  Deutsche  Zeitschr.  f.  Thiermed.,  1881  ;  Bericht  u.  d. 
Veter.-Wesen  i.  K.  Sachsen,  1885.  Karsten,  Deut.  Med.  Woch..  1884.  Mag- 
nussen,  Beitrage  zur  Diagnostik  u.  Casuistik  der  Actinomykose  :  Diss.  Kiel., 
1H85.  Mitteldorpf,  Deut.  Med.  Woch.,  1884.  Malcolm  Morris,  Brit.  Med.  Journ., 
1896.  Murphy,  New  York  Med.  Journ.,  1885.  O'Neill,  Lancet,  1886.  Pflug, 
Centralbl.  f.  d.  Med.  Wiss.,  1882.  Penfick,  Breslauer  Aerztl.  Zeitschr.,  1885  ; 
Die  Actinomykose:  Berlin,  1887.  Pusch,  Arcb.  f.  Wiss.  u.  Pr.  Thierheilk.. 
1883.  Eansome,  Brit.  Med.  Journ..  1896.  Report  of  the  Board  of  Live-Stock 
Com.  for  the  State  of  Illinois,  1890.  Roser,  Deut.  Med.  Woch..  1886.  Soltmann, 
Breslauer  Aerztl.  Zeitschr.,  1885.  Treves,  Lancet,  1884.  Zemann,  AVien.  Med. 
Jahrb.,  S.  477,  1883. 

MADURA  DISEASE. 

Boyce  and  Surveyor,  Proc.  Eoy.  Soc.,  1893  ;  Trans.  Roy.  Soc..  1894  ;  Brit. 
Med.  Journ.,  1894.  Hewlett,  Lancet,  1894.  Surveyor,  Brit.  Med.  Journ..  1892. 
Vincent,  Ann.  de  1'Institut  Pasteur,  1894. 


CHAPTER    XXXI. 

GLANDERS. 

Babes,  Acad.  de.  Med.,  1888.  Baumgarten,  Centralbl.  f.  Bacteriolog.,  1888. 
Bouchard,  Capitan  and  Charrin,  Bull,  de  1'Acad.  d.  Sc.,  Nr.  51,  1882. 
Cadeac  and  Malet,  Progres  Med.,  1886,  1887  ;  Rec.  de  Med.  Vt.,  1886  ;  Oester. 
Monatschr.  f.  Thierheilk.,  1888.  Frbhner,  Rep.  d.  Thierheilk.,  1883.  Griinwald. 
Oesterr.  Monatsschr.  fur  Thierheilk.,  Nr.  4.  1884.  Hunting,  Yet.  Record.. 
.1896.  Israel,  Berl.  Klin.  Woch.,  Nr.  11.,  1883.  Kitt,  Jahresber.  d.  Miinchen. 
Thierarzneisch,  1884.  Kranzfeldt,  Central b.  f.  Bacteriolog.,  1887.  Lbffler, 
Arbeit,  a.  d.  K.  Gesundh.  Amt.,  1886.  Loffler  and  Schutz,  Deut  Med.  Woch.. 
Nr.  52,  1882.  Molkentin,  Zur  Sicherstellung  der  Diagnose  von  Rotz :  Inaug.- 
JMss.,  1.SK',.  Raskin,  Zeit.  f.  Wiss.  Mikroscop.,  1887.  Salmon,  Reports  Bureau 
of  Animal  Industry,  1887,  1888.  Smith,  Journal  of  Comp.  Med.  and  Veterin. 
Archives,  1890.  Struck,  Deut.  Med.  Woch.,  Nos.  51  u.  52,  1883.  Vulpian  and 
Bouley,  Bull,  de  1'Acad.  de  Med.,  18X3.  Wassilieff,  Deut.  Med.  Woch.,  Nr.  ll.r 
1883.  Weichselbaum,  Wiener  Med.  Woch.,  21-24.  1884. 


BIBLIOGRAPHY.  661 

CHAPTER    XXXII. 

TETANUS.      RABIES.      LOUPING-ILL. 

TETANUS. 

Carle  and  Rattone,  Studio  Sperimentale  sull'  Etiologia  del  Tetano.  Giorn. 
clella  R.  Acad.  di  Medicina  di  Torino,  1884.  Hewlett,  Brit.  Med.  Journ.,  181)5. 
Martin,  Rep.  Med.  Off.  Local  Govt.  Board  1895.  Nicolaier,  Dent.  Mrd. 
Woch.,  Xr.  52,  1884.  Rosenbach,  Archiv  f.  Klin.  Chir.,  1886.  Roux  and 
Vaillard,  Ann.  de  1'Institut  Pasteur,  1893.  Vogel,  Deut.  Med.  Woch.,  Xr.  31, 
1884. 

RABIES. 

Babes,  Les  Bacteries,  1886.  Bauer,  Munch.  Med.  Woch.,  1886.  Bert, 
Compt.  Rend.,  1882.  Colin,  Bull.  Acad.  de  Med.  Paris,  T.  10,  1881.  Dole"ris, 
Gaz.  Med.  de  Paris,  T.  3:  Tribune  Med.  Paris,  T.  14,  1881.  Dowdeswell, 
Journ.  Roy.  Micro.  Soc.  and  Lancet,  1886.  Fol,  Acad.  des  Sciences,  1884. 
Frisch,  Wien.  Med.  Woch.,  1886.  Gibier,  Compt.  Rend.,  1883.  Kerr,  Brit. 
Med.  Journ.,  1886.  Pasteur,  Compt.  Rend.,  1881,  1884,  1886  ;  Ann.  de  M.-d. 
Veterin,  1884.  Pasteur,  Chamberland,  Roux  and  Thuiller,  Compt,  Rend.,  1882. 
Percheron,  La  Rage  et  les  Experiences  de  M.  Pasteur,  1884.  Report  of  the 
English  Hydrophobia  Com.  Vignal,  Brit.  Med.  Journ..  1886. 

LOUPIXG-ILL. 

Klein,  Journ.  of  Royal  Agric.  Soc.,  1895.  M'Fadyean,  Journ.  Royal  Agric. 
Hoc.,  1895,  and  Journ.  of  Comp.  Path,  and  Bacteriology,  1895. 

CHAPTER    XXXIII. 

FOOT-ROT. 

Brown,  Journ.  Royal  Agric.  Soc..  1892.  Nott,  Journ.  Royal  Agric.  Soc.. 
1890. 

CHAPTER    XXXIY. 

FOUL-BROOD.      INFECTIOUS   DISEASE   OF   BEES   IN   ITALY.      PEBRINE. 
FLACHERIE.      INFECTIOUS   DISEASE   OF   CATERPILLARS. 

Beehamp,  Compt.  Rend.,  1867.  Cheshire  and  Cheyne,  Journ.  of  Roy.  Micros. 
Society.  Cowan,  Journ.  Royal  Agric.  Soc.,  1892.  Forbes,  Bull.  Illin.  State 
Lab.  of  Xat.  Hist.,  1886.  Klamann,  Bienenwirthschaft.  Centralbl.,  1888. 
Pasteur,  Etudes  des  maladies  des  ver  a  soie,  1870. 


CHAPTER    XXXV. 
CLASSIFICATION   AND   DESCRIPTION   OF  SPECIES. 

Acosta  and  Rossi,  Centralbl.  f.  Bact,.  1893.  Adametz,  Die  Bakt,  der  Xutz. 
u.  Trinkwasser,  1888  ;  Landwirthschaft.  Jarhb..  1890.  Afanassiew,  St.  Peters- 
burg Med.  Woch.,  1887.  Ali-Cohen,  Centr.  f.  Bact.,  Bd.  VI.,  1889.  Almquist, 


662  APPENDICES. 

Zeitschr.  f.  Hygiene.  Bd.  X.,  1891.  Alvarez,  Compt.  Rend.,  T.  cv.,  1887. 
Arloing,  Compt.  Rend.,  cvi.  and  cvii.  Arthur,  Proc.  Acad.  Nat.  Sci.  Phil.,  1836. 
Babes,  Bakt.  Untersuch.  ii.  Sept.  Proz.  des  Kindesalters,  1889  ;  Virch.  Archiv, 
Bd.  cxv.,1889  ;  Centralb.  f.  Bacteriolog.,  Bd.  IX.,  1891  ;  Progres  Med.  Roumain. 
Babes  and  Oprescu,  Ann.  de  1'Institut  Pasteur,  Vol.  v.,  1891.  Baginsky, 
Deutsche  Med.  Woch.,  1888.  Banti,  Giornale  Medico,  1888.  Beyerinck,  Bot. 
Zeitung.  Vol.  xlix.,  1891.  Bienstock,  Zeitschr.  f.  Klin.  Med..  VIII.  Billet, 
Compt  Rend.,  T.  100.,  1885.  Biondi,  Zeitschrift.  f.  Hygiene,  Bd.  II.,  1887. 
Bizzozero,  Virchow's  Archiv,  xcviii.  Bolton,  Amer.  Journ.  Med.  Sci.,  1892  ; 
Zeitsch.  f.  Hygiene,  Bd.  I.,  1886.  Bonome,  Archiv  per  le  Sci.  Med.,  Vol.  xiii., 

1890.  Booker,  Transac.   Ninth  Internat.  Med.  Congress,  Vol.  III.     Bordoni- 
Uffreduzzi,  Fortsch.  der   Med.,    1886;    Zeitschr.   f.  Hygiene,  Bd.  III.,    1888. 
Botkin,    Zeitschr.   f.   Hygiene,    Bd.   XL,    1892.       Bouchard,    Compt.    Rend.. 
T.  cviii.,  1SS9.     Bovet,  Ann.  de  Micrographie.  Vol.  iii.,  1891.      Brannan  and 
Cheeseman,  New  York  Medical  Record,   1892.     Brefeld,    Ges.  Nat.   Freunde, 
1878.      Breunig,   Bakt.    Untersuch.   d.    Trinkwassers   der   Stadt   Kiel,    1888. 
Brieger,  Berl.  Klin.  Woch.  1884.     Brouardel  and  Boutmy,  Compt.  Rend.,  T.  92. 
p.  1056.     Bujwid,   Centralb.   f.    Bacteriolog.,    1893.      Bumm,   Der  Mikr.   der 
Gon.  Schleimk.,  1887.     Burri,  U.  eim.  z.  Zuleck.  d.  Artchar  auz  Bact.  Unter- 
such. a.  Rhein  W.  Isol.  Bact.,  1893.     Burrill,  Proc.  Amer.  Assoc.  Adyanc.  Sci., 
1880.    Caldo,  Bull,  de  la  Soc.  d'Anatom.  de  Paris,  1887.     Canon  and  Pielicke, 
Berl.   Klin.   Woch..   1892.     Caspary,    Schriften  der   Physik.   Oekon.   Ges.   zu 
Konigsberg,  Bd.  15. 1874.     Cazal  and  Vaillard,  Ann.  de  1'Instit.  Pasteur.  Vol.  v.. 

1891.  Charrin,  La  Maladie  Pyocyanique,  1889.     Cheyne,  Brit.   Med.  Journ., 
1886.     Cienkowski,  Die   Gallertbild.  des  Zuckerrubensaftes,  1878 :    Zur  Mor- 
phol.   des   Bakter.,    1876.      Clado,   Bull,   de   la   Soc.   d'Anat.   de   Paris,   1887. 
Classen,  Centr.  f.  Bakt.,  Bd.  VII.,  1890.     Cohn,  Max  Schultz's  Archiv,  Bd.  III.  : 
Ueber  zwei  Neue  Beggiatoen :   Hedwigia,  1865  ;   Beitr.  z.   Biol.  d.  Pflanzen. 
Bd.    I.,   1872.      Conn,    Centralbl.   f.   Bakteriol.,    Bd.    IX.,    1891.      Dallinger, 
Journ.   of  the   Roy.   Micro.   Soc.   London,    1878.       Dallinger  and   Drysdale, 
Monthly   Microsc.   Journ.,    1875.      Dantec,   Ann.  de  1'Institut  Pasteur,   1891. 
De  Bary,  Vergl.  Morph.  und  Biol.  der  Pilze,   1884.      Demme,   Fortschr.   der 
Med.,   1888;   Verhandl.  der  V.   Congress  fur   in   Med.   in  Wiesbaden,   1886. 
Deneke,   Deutsch.   Med.  Woch.,   1885.      Dickerhoff    and   Grawitz,   Virchow's 
Archiv,   Bd.   cii.,   1885.      Dowdeswell,    Ann.    de    Micrographie,   II.      Eberth, 
Virchow's  Archiv,  Bd.  13,  1858.     Ehrenberg,  Verhandl.  der  Berl.  Acad.,  1839. 
Eidam,  Cohn's  Beitr.  zur.  Biol.  d.  Pflanzen,  1875.      Eisenberg,    Bacteriolog. 
Diagnostik.    1891.    Emmerich,    Deutsche    Med.    Woch..    1884.      Engelmann, 
Pfliiger's  Arcb.  f.  d.  Ges.  Physiol.,  Bd.  30,  S.  95,  1882.     Engler,  Bericht.  d. 
Kommission  z.  Erfolsch.  Deutsch.  Meere.  1881.     Ernst,  Zeitschr.  f.  Hygiene, 
II.     Escherich,  Die  Darnkbakterien  des  Sauglings,  1886.     Esmarch,  Centralb. 
f.  Bact.,  lss7.    Ewart,  Proceedings  of  the  Roy.  Soc.,  1874.    Falkenheim,  Arch, 
f.  Exp.  Patholog.  u.  Pharmakol.,  Bd.  19,  1885.     Fasching,  Centralb.  Wiener 
Akad.  Wiss.,  1891.    Finkler  and  Prior,  Deutsche  Med.  Woch.   1884.     Fischel, 
Zeitschr.  f.  Heilkunde,  XII.,  1891.     Fischer,  Zeitschr.  f.  Hygiene,  1887,  1888  ; 
Centralb.  f.  Bacter.,  III.  and  IV.    Fliigge,  Microorganisms  and  Disease  (New 
8yd.    Soc.).     Foutin,   Centralb.   f.  Bacteriolog.,    1890.     Forster,  Centralbl.  f. 

icter..  II.,  1887.  Frankel  and  Franke,  Knapp.  Schweiger's  Archiv.  XVII. 
Frankland,  P.  and  G.,  Zeitschr.  f.  Hygiene,  1889  ;  Proc.  Royal  Soc.,  Vol.  xlvii., 
;90.  Freudenreich,  Ann.  de  Micrographie,  Vol.  ii.,  1890,  and  Vol.  iii., 
;91.  Frick,  Virchow's  Archiv,  CXVI.  Friedreich,  Virch.  Arch.,  Bd.  30, 
i6!.  Fulles,  Zeitsd.r.  f.  Hygiene.  Gaffky,  Langenbeck-s  Archiv  f.  Chir., 

THE 

..JHSITY 

V 


BIBLIOGRAPHY.  663 

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1878.  Gessard,  De  la  Pyocyaniue  et  de  son  Microbe,  18,s2.  Gessner,  Archiv  f. 
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Neerland  Sci.,  XXV.,  1891.  Globig,  Zeitschr.  f.  Hygiene,  III.  Gombert,  Rech. 
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Journ.,  1842.  Grotenfelt,  Fortschr.  des  Med.,  Bd.  VII.,  1889.  Guillebeau,  Ann. 
de  Micrographie,  Vol.  IV.,  1892.  Giinther,  Deutsche  Med.  Woch.,  1892  ;  Arb. 
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664  APPENDICES. 

Hunk,   Virch.  Arcb.,  Bd.  22,  1861  ;  Med.  Centralbl.,  18G4.      Muntz,   Compt. 

Rend.!  T.  cxii.    Neelsen,  Beitrag.  z.  Biol.  der  Pttanzen,  III.     Neisser,  Archiv  f. 

Hygiene.  1893.    Neumann  and  Schaeffer,  Virchow's  Archiv,  Bd.  CIX.,  1887. 

Nocard,   Ann.  de  I'lnstitut  Pasteur,  1887.      Nocard  and  Mollereau,   Ann.  de 

I'lnstitut  Pasteur,  T.  1,  1887.     Oersted,  Naturhist.  Tidsskrift,  Bd.  III.,  1840. 

Okada,  Centralbl.  f.  Bacteriolog.,  Bd.  IX.,  1891.     Paltauf  and  Heider,  Med. 

Jahrbuch.,  1889.      Passet,   Zeitschr.   f.  Hygiene,  Bd.  IX.  ;  Untersuch.  u.  die 

eitr.  Phleg.  des  Wiensch.,  1885  ;  Fortschr.  der  Med.,  1888.     Pasteur,  Ann.  de 

Chim.  et  de  Phys.,  T.  64,  1862  ;  Compt.  Rend.,  1876;  LIL,   1861.     Pansini, 

Virch.  Archiv,  Bd.  cxxii..  1890.     Perdrix,  Ann.  de  1'Institut  Pasteur,  T.  v.,  1891. 

Perty,  Zur  Kentniss.  Kleinst.    Lebensform,   1852.      Pfeiffer,    Deutsche  Med. 

Woch.,  1888  :  U.  die  Bac.  Pseudotuberculose  bei  Nagethiere,  1889  ;  Zeitschr.  f. 

Hygiene,   Bd.    VI.  ;    Bd.   VIII.,    1889.       Plagge    and  Proskauer,    Zeitsch.   f . 

Hygiene,  II.     Pohl,  Centralb.  f.  Bacteriolog.,  1892.     Pommer,  Mitth.  des  Bot. 

Inst.    zu  Gratz.,  I.     Popoff,   Ann.   de   1'Institut  Pasteur,  1890.     Prazmowski, 

Untersuch.    ii.    die    Entwick.   und    Fermentwirkung  einer   Bakterien,    1880. 

Prove,  Beitr.  z.  Biolog.  der  Pflanzen,  Bd.  III.     Raczynsky,  Diss.  der  Militar. 

Med.  Akad.,  1888.     Keimann,    Inaug.    Diss.  Wurzburg,   1887.     Renon.   Ann. 

de  1'Institut  Pasteur,  1892.      Roscoe  and  Lunt,  Phil.  Trans.  Roy.  Hoc.,  1892. 

Rosenthal,  Zeitschr.  f.  Hygiene,  V.     Roth,  Zeitschr.  f.  Hygiene,  Bd.  VIII., 

1890.     Russell,    Zeitschr.    f.   Hygiene,   Bd.  XL,    1891.      Sakharoff,   Ann.   de 

I'lnstitut  Pasteur,  1891.     Sanarelli,  Centralbl.  Bd.  IX.  1891.      Scheibenzuber, 

Allgem.   Wien.    Med.   Zeit.,   1889.      Scheurlen,   Deutsche   Med.  Woch,  1888. 

Schimmelbusch,  Deut,  Med.  Woch.,  1889.      Scholl,  Fortschr.    der   Med.,  VII. 

Schottelius,    Biol.   Untersuch.  ii.  dem   Micrococcus   Prodig.,  1887  ;   Zeitschr. 

f.   Hygiene,    1890.      Schrb'ter,   Beitrage   z.   Biol.   d.    Pflanz.,     Bd.    1,    H.    2. 

Schutz,  Archiv  f.  Wiss.  und  Prakt.   Thierheilk.,  1888.      Senger,   Berl.   Klin. 

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de  Microscop.,  1888.  Van  Zieghem,  Compt.  Rend.,  Ixxxviii.  and  Ixxxix.,  1879. 
Vaughan,  Amer.  Journ.  Med.  Sci.  1892.  Vignal,  Archiv  de  Phys.,  viii.,  1886  ; 
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Bd.  I.  Warington,  Journ.  of  Chemical  Soc.,  1890.  Weibel,  Central,  f.  Bak- 
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Ziegler's  Beitrage,  Bd.  IV.,  1888  ;  Das  Osterreich.  Samtalswesen,  1889.  Weisse, 
Bull.  Phys.-Mathemat.  de  St.  Petersbourg,  III.,  1845.  Weisser,  Zeitsch.  f. 
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BIBLIOGRAPHY.  665 

APPENDICES. 

ELEMATOZOA. 

Bardels.  Gaz.  des  Hopit..  1882.  Crookshank,  Journ.  Eoy.  Micr.  Soc.. 
1886.  Cuboni  and  Marchiafava,  Arch.  f.  Exp.  Pathol.,  Bd.  13,  1880  ;  Atti  della 
R.  Acad.  del  Lincei.,  1881.  Danilewsky,  La  Parasitologie  Comparee  du  Sang, 
1889;  Centr.  f.  Bakt.  u.  Parasitenk.,  1891.  Gerhardt,  Zeitschr.  f.  Klin.  Med., 
P,d.  7,  1884.  Golgi,  Archivio  per  le  Scienze  Mediche,  1886.  Grass!  and 
Feletti,  Centr.  f.  Bakt.  u.  Parasitenk.,  1891.  Klebs  and  Tommasi-Crudeli,  Arch, 
f .  Exp.  Pathol..  Bd.  2, 1879.  Kotelmaun,  Virchow's  Arch.,  Bd.  97, 1884.  Laveran, 
Compt.  Kend.,  1881  ;  Ibid.,  No.  17,  1882  ;  Traite  des  Fievres  Palustres,  1884. 
Xeoni,  Gazetta  Medica  di  Roma.  1*84.  Mannaberg,  Trans.  New  Syd.  Soc. 

1894.  Manson,   Brit.  Med.  Journ.,  189(5.     Marchand,  Virch.  Archiv,  Bd.  88, 
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1892;  Journ.  of  Comp.  Path.,  Vol.  I.  Bussell,  Brit.  Med.  Journ.,  1890.' 
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1895.  Soudakewitch,  Ann.  de  1'Institut  Pasteur,  1892.    Wickham,  Archiv.  de 
Med.  Exper.,  1890. 


SUPPLEMENTARY    APPENDIX. 


EXTRACTS  FROM  THE  FINAL  REPORT  OF  THE  ROYAL 
VACCINATION   COMMISSION. 


THE  final  Report  of  the  Royal  Commission  ito  inquire  into  the 
subject  of  Vaccination  was  published  on  September  18th,  1896.* 
The  author  desires  to  gratefully  acknowledge  the  permission  granted 
him,  by  the  Controller  of  Her  Majesty's*  Stationery  Office,  to  make 
extracts  bearing  more  especially  on  the  history  and  pathology  of 
protective  inoculation  and  the  prevention  of  small-pox.  The  reader 
is  recommended  by  the  author  to  study  the  whole  of  the  report. 


History  of  S 

The  early  history  of  small-pox,  like  that  of  many  similar  diseases,  is 
obscure,  is  subject  to  much  debate,  and,  save  perhaps  on  one  point,  is  of 
antiquarian  interest  only. 

The  records  of  the  eighteenth  century  show  that  the  disease  was  very 
prevalent  in  western  Europe  during  the  whole  of  that  century.  The 
records  of  the  seventeenth  century  also  show  that  small-pox  was  a  very 
common  disease  during  that  century  :  this  is  especially  the  case  as  regards 
the  latter  half  of  the  century.  The  statistics  which  exist  with  respect  to 
Geneva,  and  various  scattered  statements,  further  show  that  small-pox 
was  a  well-known  disease  in  the  sixteenth  century  ;  but,  except  for  the 
records  which  are  said  to  exist  of  severe  epidemics  in  Iceland  taking 
place  as  early  as  1241,  as  we  go  further  back  the  evidence  as  to  the 
existence  of  the  disease  becomes  less  and  less  clear,  and  indeed  debate  - 
able,  depending  as  it  does  largely  on  the  interpretation  of  incidental 

*  The  report  may  be  obtained  either  directly,  or  through  any  bookseller, 
from  Eyre  &  Spottiswoode,  East  Harding  Street,  Fleet  Street,  B.C.,  and  32, 
Abingdon  Street,  Westminster,  S.W.  ;  or  John  Menzies  &  Co.,  12,  Hanover 
Street,  Edinburgh,  and  90,  West  Nile  Street,  Glasgow  ;  or  Hodges.  Figgis  & 
Co.,  Limited,  104,  Grafton  Street,  Dublin. 

f  The  heading  to  the  extracts  from  the  Report  of  the  Commission  are  mine. 
—  E.  M.  C. 

067 


668  SUPPLEMENTARY  APPENDIX. 

statements  in  various  medical  and  other  writings.  There  seems,  how- 
ever, to  be  adequate  proof  of  the  prevalence  of  small- pox  in  the  East, 
in  Asia  Minor  and  other  countries,  even  in  the  earlier  centuries  of  the 
Christian  era. 

A  view  very  generally  taken  teaches  that  small-pox,  introduced  from 
the  East,  began  to  be  common  in  western  Europe  during  the  fifteenth 
century,  though  perhaps  existing  still  earlier;  that  it  increased  during 
the  sixteenth  and  seventeenth  centuries,  especially  the  latter  ;  and  that  it 
was  very  prevalent  during  the  eighteenth  century. 

In  dealing  with  the  eighteenth  century  it  must  be  borne  in  mind  that 
during  the  second  half  of  the  century  the  natural  conduct  of  small-pox, 
as  we  shall  see  later  on,  was  modified  by  the  practice  of  inoculation— that 
is,  by  the  artificial  giving  of  the  disease  by  the  introduction  of  the  virus 
through  a  wound  in  the  skin. 

Our  knowledge  of  the  history  of  small-pox  in  western  Europe  during 
the  seventeenth  and  eighteenth  centuries  is  very  largely  based  on  the 
official  records  known  as  the  "  London  Bills  of  Mortality."  Official 
records  bearing  on  small-pox-  are  furnished  by  Geneva,  going  back  as 
far  as  the  sixteenth  century,  by  Sweden,  going  back  to  the  year  1749, 
and  by  some  other  places.  Data  are  also  furnished,  especially  for  the 
latter  part  of  the  eighteenth  century,  by  parish  records  in  various  parts 
of  Great  Britain  reaching  over  a  variable  number  of  years,  as  well  as  by 
scattered  statements  in  various  works. 

These  Bills  of  Mortality  form  by  far  the  most  complete  source  of  our 
knowledge  of  small-pox  in  England  in  past  times  ;  but  it  must  be  borne 
in  mind  that  in  respect  to  any  contagious  disease  like  small-pox  the 
conditions  of  London  were  peculiar.  The  population  was  to  a  marked 
extent  a  moving  one  ;  a  large  number  of  persons  were  continually 
entering  London  or  leaving  it,  were  passing  to  and  from  it,  from  and  to 
the  provinces  of  England  and  other  countries.  Of  these  persons,  some, 
coming  from  infected  districts,  brought  into  London  fresh  sources  of 
contagion  ;  others  again,  coming  from  districts  free  from  small-pox,  and 
never  having  had  the  disease,  brought  into  London  fresh  material  to  serve 
as  food  for  the  disease.  Further,  London  presented  in  an  exaggerated 
degree  the  two  features  of  a  great  city  which  have  a  great  influence  on 
the  progress  and  characters  of  a  contagious  disease  like  small-pox.  The 
crowding  both  of  the  dwelling-places  and  the  thoroughfares,  as  well  as 
the  movement  continually  going  on,  multiplied  the  opportunities  for  the 
spread  of  disease,  and  the  accompanying  insanitary  conditions,  as  well  as 
the  greater  inducement  to  irregular  living,  tended  to  increase  the  severity 
of  the  disease  when  taken,  and  to  heighten  the  mortality  from  it.  The 
history  of  small-pox  in  London  must  not  be  taken  as  representative  of 
the  history  of  small-pox  in  England  generally. 

Inoculation    of   Small-pox. 

The  practice  of  inoculation  for  the  small-pox— that  is,  the  artificial 
introduction  of  the  virus  into  the  system  by  the  insertion  of  fluid  from 
a  vanolous  pustule  into  wounds  of  the  skin  made  for  the  purpose  - 
began  definitely  m  England  towards  the  end  of  the  first  quarter  of  the 


REPORT   OF   THE   ROYAL    VACCINATION   COMMISSION.  669 

eighteenth  century.  Attention  was  directed  to  the  matter  by  letters 
from  Timoni  of  Athens  (dated  1713)  and  Pylarini,  published  in  the 
29th  volume  of  the  Philosophical  Transactions  (1716),  and  especially 
by  a  letter  from  Lady  Mary  Wortley  Montagu  in  1717.  Though 
there  are  indications  that  in  Great  Britain  and  Ireland,  as  in  other 
countries,  some  sort  of  inoculation  had  occasionally  been  practised  at 
a  much  earlier  date,  the  first  clearly  recorded  case  in  England  is  that 
of  the  daughter  of  Lady  Mary  Wortley  Montagu  (whose  son  had  some 
time  before  been  inoculated  at  Constantinople),  inoculated  by  Maitland, 
in  London,  in  April  1721.  Other  cases  soon  followed  in  England,  and 
about  the  same  time  the  practice  was  also  introduced  in  other  countries 
of  western  Europe,  and  into  the  United  States  of  America,  namely,  at 
Boston. 

It  was  found  that  the  attacks  induced  by  inoculation  were  as  a  rule 
milder,  and  very  much  less  fatal,  than  the  attacks  of  the  "natural'' 
disease,  the  fever  and  constitutional  disturbance  being  less  and  of  shorter 
duration,  and  the  eruptive  pustules  much  fewer  :  the  number  of  these 
varied,  being  commonly  a  dozen  or  two,  somitines  only  two  or  three, 
sometimes  a  hundred  or  more.  In  som  i  cases  there  was  no  eruption 
at  all,  the  effect  being  limited  to  constitutional  disturbances  and  to 
changes  in  the  wounds  of  inoculation  themselves  ;  it  was  maintained  that 
in  such  cases  the  disease  had  really  been  taken,  and  immunity  against 
a  subsequent  attack  secured,  as  in  case?  of  natural  small-pox  or  of 
inoculated  small-pox  manifesting  itself  in  an  eruption  of  pustules. 

In  England  the  practice  of  inoculation  at  its  introduction,  though 
much  lauded  and  strongly  urged  by  some,  was  bitterly  opposed  by  others. 
Moreover,  the  initial  enthusiasm  in  favour  of  it  soon  declined,  so  that 
in  the  years  1730-40  very  little  inoculation  seems  to  have  been  practised. 
About  1740,  however,  a  revival  appears  to  have  taken  place :  in  1746 
an  Inoculation  and  Small-pox  Hospital  was  started  in  London  ;  and 
during  the  whole  of  the  latter  half  of  the  eighteenth  century  the  practice 
may  be  said  to  have  been  very  general.  It  was  especially  so  during  the 
last  quarter  of  the  century,  the  increase  being  at  least  largely  due 
to  the  "improved  methods"  of  inoculation  introduced  by  one  Sutton 
in  1763,  and  known  as  "  the  Suttonian  method." 

Since  an  inoculated  person  was  infectious,  each  inoculation  was 
a  source  of  danger  to  those,  not  protected  by  a  previous  attack,  who 
came  into  the  company  of,  or  even  near,  the  inoculated  person  during 
the  attack  ;  and  this  danger  was  increased  by  the  fact  that  the  mild 
character  of  the  inoculated  disease  permitted,  in  many  cases  at  least, 
the  patient  to  move  about  among  his  fellows.  Moreover,  as  Haygarth, 
himself  a  zealous  advocate  of  inoculation  in  a  systematic  regulated 
manner,  points  out,  the  beneficial  results  of  inoculation  had  robbed 
the  disease  of  its  terrors  to  so  great  an  extent  that  the  rich  and  powerful 
no  longer  made  the  efforts  which  they  formerly  did  to  prevent  its 
entrance  into,  or  its  spread  in,  their  neighbourhood,  and  thus  favoured 
its  spread  among  the  unprotected  poor  ;  so  that  inoculation  "  though 
eminently  useful  to  the  rich  appeared  to  be  injurious  to  the  poor." 
Adding,  therefore,  together  the  cases  of  inoculated  small-pox,  and  the 


670  SUPPLEMENTARY  APPENDIX. 

cases  of  natural  small-pox  of  which  the  inoculated  cases  were  in  one  way 
or  other  the  cause,  it  seems  probable  that  inoculation  did  tend  to  increase 
the  prevalence  of  small-pox  ;  but  there  are  no  recorded  data  to  show  that 
this  really  was  the  case,  and  this  supposed  influence  may  have  been 
counterbalanced  by  other  influences. 

The  evidence  as  to  the  influence  which  inoculation  had  on  the 
mortality  from  small-pox  is  in  many  respects  conflicting.  Haygarth, 
though  he  admits  that  in  other  parts  of  the  kingdom  the  practice  may 
have  saved  many  lives,  was  persuaded  that  in  his  own  part  of  England 
and  Wales  the  deaths  by  the  small-pox  had  been  augmented  by  it  ;  and 
he  points  out  that  in  London,  Geneva,  and  other  "  towns  in  different 
situations  and  circumstances,  the  mortality  from  this  distemper  has 
increased  since  the  introduction  of  inoculation."  Several  writers  in  the 
latter  part  of  the  last,  and  the  early  part  of  the  present  century,  held 
a  similar  view.  Other  writers,  again,  opposed  this  view. 

Tradition  of  the  Dairy-folk. 

There  was  at  the  close  of  the  eighteenth  century,  if  not  earlier,  in 
districts  where  cow-pox  had  appeared,  a  belief  among  the  dairy-folk  that 
those  who  had  taken  the  cow-pox  never  took  the  small-pox  ;  and  indeed 
one  Jesty,  a  Dorsetshire  farmer,  had  in  1774,  in  the  case  of  his  wife  and 
sons,  purposely  introduced  the  matter  of  cow-pox  into  the  human  subject 
with  the  view  of  protecting  from  small-pox. 

Cow-pox. 

Vaccinia  or  cow-pox  is  a  disease  affecting  milch  cows,  and  marked  by 
an  eruption  on  the  udder  and  teats.  The  disease  can  be  communicated 
from  the  cow  to  man.  Dairymen  and  maids  engaged  in  milking  cows 
affected  with  cow-pox  are  apt  to  have  sores  of  a  special  kind  on  their  hands 
or  elsewhere,  the  development  of  the  sores  being  frequently  accompanied 
by  febrile  symptoms.  There  can  be  no  doubt  that,  in  a  certain  number 
of  cases  at  all  events,  such  sores  are  the  local  manifestations  of  cow-pox  : 
the  virus  from  the  eruption  on  the  cow  being  introduced  into  some  scratch 
or  other  imperfection  in  the  skin  of  the  milker  and  there  producing  its 
local  effects,  accompanied  more  or  less  by  general  symptoms. 

Inoculation  of  Cow-pox. 

The  practice,  however,  of  inoculating  with  the  matter  of  cow-pox,  or 
vaccination  as  it  was  subsequently  called,  may  be  considered  as  dating 
from  the  publication  of  the  "  Inquiry  into  the  Causes  and  Effects  of  the 
Variolae  Vaccine  "  of  Edward  Jenner,  published  in  the  summer  of  the 
year  1798.  The  practice  rapidly  spread,  and  prevailed  widely  in  this 
country  and  other  parts  of  western  Europe  during  the  first  quarter  of  the 
present  century.  It  was,  beyond  all  question,  so  adopted  in  the  genuine 
belief  that  it  afforded  protection  against  small-pox. 

In  the  treatise  to  which  reference  has  been  made  Jenner  records  in  the 
first  place  a  number  (19)  of  cases  in  which  a  person  who  had  accidentally 
taken  cow-pox  from  the  cow  had  never  had  small-pox,  and  appeared 


REPORT   OF  THE   ROYAL   VACCINATION   COMMISSION.  671 

incapable  of  taking  that  disease  ;  the  insusceptibility  being  shown  on  the 
one  hand  by  the  failure  to  contract  the  disease  after  ample  exposure  to 
contagion,  such  as  nursing  and  attending  to  or  even  sleeping  with  persons 
suffering  from  small-pox,  and  on  the  other  hand  by  the  fact  that  when 
the  person  in  question  was  inoculated  with  the  matter  of  small-pox  in  the 
manner  then  usual  (the  matter  being  tested  as  to  its  efficiency  on 
susceptible  persons)  the  inoculation  failed  to  excite  small-pox.  In  the 
course  of  the  inoculation  practice  it  had  been  observed  that  when  the 
operation  was  performed  upon  a  person  who  had  already  had  small-pox, 
either  naturally  or  by  inoculation,  the  wound  of  inoculation,  instead  of 
developing,  as  it  did  when  the  operation  was  successful  in  a  person  who 
had  not  had  the  small-pox,  into  a  vesicle  and  so  into  a  pustule  with  the 
variolous  characters  (the  development  being  accompanied  by  febrile 
symptoms  and,  save  in  exceptional  cases,  by  the  appearance  of  a  smaller 
or  greater  number  of  variolous  pustules  on  parts  of  the  skin  other  than 
the  seat  of  inoculation),  presented  as  a  rule  nothing  more  than  some  slight 
inflammation,  dying  away  in  a  few  days  without  any  other  symptom,  or  even 
healed  at  once  without  any  symptoms  at  all,  local  or  general  ;  and  in  the 
exceptional  cases  in  which  further  changes  took  place  in  the  wound,  these 
were  not  accompanied  or  followed  by  an  eruption  of  pustules  or  even  by 
the  febrile  and  other  general  symptoms  of  small- pox.  Accordingly,  in 
cases  of  small-pox  inoculation  where  it  was  doubtful  whether  the  disease 
had  been  communicated,  it  had  become  not  an  uncommon  practice  to 
repeat  the  operation,  in  order  to  judge  by  the  effects  produced  whether 
the  earlier  inoculation  had  or  had  not  produced  the  disease  ;  and  the 
practice,  thus  originating  in  connexion  with  small-pox  inoculation,  had 
come  to  be  spoken  of  as  the  "  variolous  test." 

In  his  treatise  Jenner  distinguishes  between  what  he  calls  true  cow-pox 
and  other  eruptions  which  he  speaks  of  as  spurious,  and  which  he  regarded 
as  not  affording  protection  against  small-pox,  although  he  gives  no  details 
to  show  that  the  cases  quoted  by  him  as  affording  protection  were  cases 
of  his  true  cow-pox.  He  also  developed  the  view  that  matter  derived  from 
horses  suffering  from  the  disease  known  as  the  grease  is  capable  of  giving 
rise  to  cow-pox  in  the  cow,  and  indeed  is  the  real  origin  of  the  true 
disease.  It  may  be  added  that  Jenner  also  expressed  the  opinion  that  the 
protection  thus  afforded  by  cow-pox  was  permanent  in  character. 

Jenner  further  recorded  in  the  same  treatise  how  he  had  in  17% 
inoculated  a  healthy  boy  of  eight  years  of  age  in  the  arm  with  cow-pox 
matter  taken  from  a  sore  on  the  hand  of  a  dairymaid  who  had  been 
infected  with  the  disease  by  milking  cows  suffering  from  cow-pox.  He 
describes  the  appearances  subsequently  presented  by  the  wounds,  and 
states  that,  six  weeks  afterwards,  the  results  of  inoculating  the  boy  with 
variolous  matter  were  those  commonly  seen  to  follow  the  inoculation  of 
persons  who  had  previously  had  the  cow-pox  or  the  small-pox  :  that  is 
to  say,  the  "  variolous  test"  showed  the  boy  to  be  insusceptible  to  small- 
pox. Some  months  afterwards  the  boy  was  again  inoculated,  but  no 
sensible  effect  was  produced  on  the  constitution.  Jenner  then  relates  that 
subsequently,  in  the  spring  of  1798,  he  inoculated  a  child,  and  obtained  a 
similar  result  with  matter  taken  directly  from  the  nipple  of  a  cow  infected 


672  SUPPLEMENTARY  APPENDIX. 

with  cow-pox  ;  from  the  pustule  on  the  arm  of  this  child  he  inoculated 
another,  and  from  this  again  several,  and  from  one  of  these  latter  a  fourth 
in  succession,  and  then  a  fifth.  To  three  of  these  the  "  variolous  test " 
was  applied,  and  it  is  stated  with  the  same  results. 

Woodville's  Lymph. 

The  experiences  of  Jenner  did  not  stand  alone.  His  results  and 
views  attracted  great  attention,  and  in  the  early  part  of  the  year  1799 
Woodville  and  Pearson,  who  were  physicians  to  the  Small-pox  Hospital 
in  London,  commenced  making  experiments  with  vaccine  matter  with 
a  view  to  ascertain  whether  it  afforded  protection  against  small-pox. 
They  arrived,  like  Jenner,  at  the  conclusion  that  it  did. 

In  January  1799  Woodville,  having  found  cow-pox  to  be  present  in 
a  "  dairy  "  at  Gray's  Inn  Lane,  inoculated  seven  persons  at  the  Small- 
pox Hospital  with  matter  from  one  of  the  cows  at  the  "dairy,"  and 
other  persons  with  matter  from  sores  on  a  dairymaid  employed  at 
the  same  place  who  had  become  infected  from  the  cows.  From  these 
cases  he  inoculated  in  succession  others  at  the  Hospital,  eventually  to 
the  number  of  many  hundreds,  and  thus  established  the  stock  of  what 
has  been  spoken  of  as  "  Woodville's  lymph."  Pearson  also  at  the  same 
time  occupied  himself  with  the  question  of  inoculation  with  the 
cow-pox,  writing  a  pamphlet  about  it.  Woodville  and  he  distributed 
to  many  persons  in  this  country  and  abroad  quantities  of  the  lymph 
from  the  Hospital  ;  and  this  was  the  beginning  of  the  more  general 
practice  of  vaccination,  for  Jenner's  stock  of  lymph,  the  results  of 
which  he  had  described  in  his  treatise,  had  come  to  an  end. 

Although  Woodville's  "Hospital  lymph"  appears  to  have  been 
widely  distributed  by  himself  and  by  Pearson,  and  thus  to  have  been 
the  source  of  the  lymph  used  in  vaiious  places  in  the  early  days  of 
vaccination,  it  was  not  the  only  source,  even  in  those  days.  Pearson 
also  obtained  lymph  from  cow-pox  at  a  dairy  in  the  Marylebone  Road, 
and  used  this  "  in  certain  situations,"  which  may  be  presumed  to 
include  places  elsewhere  than  in  the  Hospital.  He  also  speaks  of 
having  obtained  lymph  from  the  cow  from  a  third  source.  Jenner 
again,  who  received  and  used  some  of  Woodville's  Hospital  lymph,  also 
obtained  lymph  from  some  other  sources  :  for  instance,  from  a  cow 
at  a  Mr.  Clark's  farm  in  Kentish  Town.  Further,  Woodville  in  1800 
speaks  of  his  having  at  various  times  procured  the  vaccine  virus  as 
produced  in  different  cows,  which  when  used  at  the  Hospital  produced 
the  same  effects  as  the  Gray's  Inn  Lane  lymph.  We  are  not  justified 
in  assuming  that  an  account  of  every  new  source  of  lymph  was 
published  ;  and  there  may  have  bsen  others,  it  is  impossible  to  say 
how  many,  than  those  just  mentioned.  In  any  case  Woodville'.s 
Hospital  lymph  was  not  the  only  lymph  used  in  those  early  days  ;  not 
only,  however,  was  it  largely  used  (indeed,  we  have  no  evidence  of  so 
widespread  a  use  of  lymph  derived  from  any  other  source),  but  the 
u?e  of  it  marks  the  definite  beginning  of  the  practice  of  vaccination  ; 
and  the  history  of  it  demands  special  notice. 


REPORT   OF   THE   ROYAL   VACCINATION   COMMISSION.  673 

Of  the  cases  recorded  by  Woodville  in  his  Reports,  the  larger 
number  (about  three-fifths)ipresented  an  important,  and,  as  compared  with 
Jenner's  cases,  a  new  feature,  in  that,  in  addition  to  the  changes  taking 
place  at  the  seat  of  inoculation  and  constituting  what  Woodville  called 
the  "cow-pox  tumour,"  which  may  here  be  spoken  of  as  the  "vaccine 
vesicle,"  an  eruption  over  the  body  of  a  greater  or  less  number  of 
pustules  was  observed.  These  eruptive  pustules  occurred  in  the  very 
first  cases  :  of  the  seven  cases  inoculated  from  the  cow,  four,  and  of 
the  five  inoculated  from  the  dairymaid,  four,  had  such  pustules  ;  and 
their  appearance  is  recorded  again  and  again  in  the  series,  down  to  the 
case  which  appears  last  but  one  in  the  tabular  statement  forming  part 
of  the  Reports. 

Moreover,  an  eruption  of  pustules  is  described  in  certain  of  the  cases 
of  which  accounts  were  published  at  about  the  same  time  by  Pearson 
and  many  others.  In  some  of  these  cases  the  lymph  used  was  supplied 
from  the  Small-pox  Hospital  by  Woodville  or  Pearson. 

It  must  be  admitted  that  these  pustules  were  pustules  of  small-pox,, 
and  that,  therefore,  Woodville's  cases,  which  did  so  much  to  establish 
the  practice  of  vaccination,  were  not  cases  simply  of  cow-pox  but  of 
cow-pox  mixed,  so  to  speak,  with  small- pox.  It  has  indeed  been 
maintained  that  Woodville's  cases  were  not  cases  of  cow-pox  at  all — 
that  small-pox  was  inadvertently  introduced  into  the  very  first  cases ; 
that  the  history  of  the  whole  series  is  the  history  of  a  series  of  small-pox 
cases  putting  on  special  characters,  and  that  therefore  the  lymph  used 
and  distributed  by  Woodville  and  Pearson  was  in  reality  not  cow-pox 
lymph  but  small-pox  lymph.  A  review  of  all  the  evidence  available 
leads  to  no  other  conclusion  than  that,  however  much  in  Woodville's, 
Pearson's  and  other  cases  cow-pox  was  mixed  up  with  small-pox,  the 
lymph  used  and  distributed  by  Woodville  and  Pearson  and  called  by 
them  cow-pox  lymph  (excluding  of  course  all  the  cases,  of  which  there 
were  not  a  few,  in  which  matter  was  taken  not  from  the  local  ;;  cow-pox 
tumour"  at  the  seat  of  inoculation,  but  from  one  of  the  eruptive 
pustules)  was  veritable  cow-pox  lymph  having  the  true  characters  of 
cow-pox  lymph  only. 

It  of  course  follows  that  the  cases,  both  in  Woodville's  practice  and 
in  that  of  others,  in  which  the  inoculation  of  cow-pox  matter  wa& 
accompanied  by  an  eruption  of  pustules,  due  to  small-pox  being  present 
as  well  as  cow-pox,  when  appealed  to  as  showing  immunity  against 
small-pox  (by  the  test  either  of  exposure  to  contagion  or  of  inoculation), 
furnished  false  evidence  as  to  that  immunity  being  due  to  cow-pox  ;. 
it  might  have  been  due  to  the  accompanying  small-pox.  So  far  then  as 
the  adoption  of  vaccination  was  assisted  by  cases  of  this  description,  it 
may  be  held  to  have  rested' on  erroneous  data. 

The  Decline  of  Small-pox. 

One  effect  of  the  introduction  of  vaccination  was  a  very  great  decrease 
in  the  practice  of  inoculation,  which  had  become  very  prevalent  during  the 
later  part  of  the  previous  century.  And  the  view  has  been  put  forward. 

43 


674  SUPPLEMENTARY   APPENDIX. 

that,  the  prevalence  of  inoculation  having  greatly  increased  the  amount 
of  small-pox,  the  diminution  of  small-pox  in  question  was  the  result  of 
the  decrease  of  inoculation. 

The  question  how  far  the  behaviour  of  small-pox  in  the  eighteenth 
century  and  earlier  was  influenced  by  sanitary  conditions,  is  one  rendered 
difficult  by  the  lack  of  exact  information.  We  may  distinguish  between 
overcrowding  as  one  insanitary  condition  and  all  other  insanitary  condi- 
tions, such  as  lack  of  cleanliness  and  the  like.  A  priori  we  should  ex- 
pect that  a  dense  population,  especially  one  of  great  internal  movement, 
and  one  in  continual  interchange  with  surrounding  populations,  by 
offering  greater  facilities  for  the  conveyance  of  contagion,  would  lead  to 
a  greater  amount  of  small-pox.  London  was  a  conspicuous  instance  of 
the  above,  and  the  apparent  greater  prevalence  of  small-pox  in  London 
than  in  the  provinces  may  be  attributed  to  these  causes  :  but  it  would 
appear  that  the  increase  was  felt — as  indeed  would,  a  priori,  seem  pro- 
bable— rather  in  the  constant  presence  of  small-pox  to  a  considerable 
amount  at  all  times  than  in  the  mortality  of  the  epidemics  when  these 
occurred.  And  the  same  seems  also  to  be  shown  to  a  less  extent  in  other 
large  cities,  such  as  Liverpool.  But  in  this  matter  of  dense  and  moving 
populations  the  eighteenth  century  did  not  differ  markedly  from  the 
early  part  of  the  nineteenth.  We  might  a  priori  expect  the  other 
acknowledged  imperfect  sanitary  conditions  of  the  eighteenth  century  to 
increase  the  fatality  of,  and  so  to  a  corresponding  extent  the  mortality 
from,  small-pox  ;  but  there  is  no  exact  evidence  to  confirm  this  supposi- 
tion. If  on  the  contrary  we  recognise  that  in  the  course  of  the  eighteenth 
century  the  general  mortality,  the  relative  number  of  deaths  from  all 
causes,  went  on  decreasing,  and  attribute,  as  has  been  done,  this  decrease 
to  improved  sanitary  conditions,  no  like  decrease  of  small-pox  took  place. 
Again,  the  places  which  were  deemed  the  most  salubrious  appear  to  have 
been  visited  by  epidemics  of  small-pox  as  severe  as  those  which  fell  on 
unhealthy  places.  Thus  the  epidemic  in  Chester  in  1774  was  undoubtedly 
a  severe  one,  and  yet  Haygarth  writes,  "  The  healthiness  of  Chester,"  as 
shown  by  statistics,  "  must  appear  so  very  extraordinary  as  to  be  almost 
incredible."  And  in  general  both  the  incidence  of,  and  mortality  from, 
small-pox  seem  to  have  been  far  less  affected  by  sanitary  conditions  than 
might  a  priori  have  been  expected. 

It  may  be  urged  against  the  view  that  the  decline  of  small-pox  was 
due  to  improved  sanitary  conditions,  in  the  first  place,  that,  admitting 
the  introduction  of  sanitary  improvements,  no  evidence  is  forthcoming 
to  show  that  during  the  first  quarter  of  the  nineteenth  century  these 
improvements  differentiated  that  quarter  from  the  last  quarter,  or  half, 
of  the  preceding  century  in  any  way  at  all  comparable  to  the  extent 
of  the  differentiation  in  respect  to  small- pox.  In  the  second  place, 
admitting  a  priori  that  crowded  dwellings  tend  to  increase  the  liability  to 
contagion,  and  so  the  prevalence  of  the  disease,  while  other  insanitary 
conditions  tend  in  addition  to  increase  the  fatality  among  those  attacked, 
that  insanitary  conditions  as  a  whole  must  tend  to  increase  the 
mortality  from  small-pox,— no  evidence  is  forthcoming  which  distinctly 
shows  that  the  dependence  of  the  prevalence  of,  or  the  mortality  from, 


REPORT   OF  THE   ROYAL   VACCINATION   COMMISSION.  675 

small-pox,  on  the  lack  of  sanitary  conditions,  was  a  feature  of  the  history 
of  small-pox  during  the  eighteenth  century. 

Upon  the  whole,  then,  we  think  that  the  marked  decline  of  small-pox 
mortality  in  the  first  quarter  of  the  present  century  affords  substantial 
evidence  in  favour  of  the  protective  influence  of  vaccination. 

Age  Incidence  of  Small-pox. 

A  study  of  the  age  incidence  of  small-pox  mortality  is  very  instructive. 
In  connexion  with  this  point  it  is  necessary  to  bear  in  mind  that  experi- 
ence has  led  to  the  conclusion  that,  whatever  be  the  protective  effect  of 
vaccination,  it  is  not  absolutely  permanent  ;  the  most  convinced  advocates 
of  the  practice  admit  that  after  the  lapse  of  nine  or  ten  years  from  the 
date  of  the  operation  its  protective  effect  against  an  attack  of  small-pox 
rapidly  diminishes,  and  that  it  is  only  during  this  period  that  its  power  in 
that  respect  is  very  great ;  though  it  is  maintained  that,  so  far  as  regards 
its  power  to  modify  the  character  of  the  disease  and  render  it  less  fatal, 
its  effect  remains  in  full  force  for  a  longer  period,  and  never  altogether 
ceases.  The  experience  upon  which  this  view  is  founded  is  derived  almost 
exclusively  from  the  case  of  infantile  vaccination.  It  has  been  supposed 
by  some  that  the  transitory  character  of  the  protection  results  from 
changes  connected  with  the  growth  from  infancy  to  adult  years.  Whether 
this  be  so  or  not,  we  have  no  means  of  determining. 

No  doubt,  when  Jenner  drew  the  attention  of  the  public  to  the  value 
of  vaccination,  he  believed  that  a  single  successful  inoculation  of  vaccine 
matter  secured  absolute  immunity  for  the  future  from  an  attack  of  small- 
pox. It  is  certain  that  in  this  he  was  mistaken.  It  may  well  be  doubted 
whether  the  anticipation  was  a  reasonable  one.  No  such  immunity  is 
secured  by  an  attack  of  small-pox,  though  there  are  few  who  would 
maintain  the  proposition  that  it  is  without  protective  influence  against 
another  attack.  A  priori  there  would  seem  to  be  no  sound  ground  for 
expecting  that  vaccinia  would  afford  more  potent  protection  than  small- 
pox itself.  The  extent  of  the  protection  afforded  (assuming  that  there  is 
some  protective  influence)  could  only  be  determined  by  experience.  It  soon 
became  apparent  that  Jenner  had,  in  the  first  instance,  overrated  the 
effect  of  vaccination.  That  he  should  thus  have  overestimated  it 
is  not  to  be  wondered  at,  when  the  tendency  to  be  unduly  sanguine, 
which  besets  the  discoverer  of  any  new  prophylactic,  and,  indeed,  every 
discoverer,  is  borne  in  mind. 

We  think,  taking  it  all  together,  that  the  evidence  bearing  upon 
the  question  whether  the  vaccinated  are  less  liable  to  be  attacked  by 
small-pox  than  the  unvaccinated,  points  to  two  conclusions  :  first,  that 
there  is,  taking  all  ages  together,  less  liability  to  attack  among  the 
vaccinated  than  among  the  unvaccinated  ;  and  next,  that  the  advantage  in 
this  respect  enjoyed  by  vaccinated  children  under  ten  years  of  age  is 
greatly  in  excess  of  that  enjoyed  at  a  more  advanced  period  of  life. 

In  considering  whether  vaccination  has  been  the  principal  cause  of  the 
decline,  we  must  inquire  whether  the  other  causes  suggested  by  those  who 
deny  the  efficacy  of  vaccination  will  satisfactorily  account  for  it. 


676  SUPPLEMENTARY    APPENDIX. 

Effect  of  Sanitation. 

It  is  said  that  the  decline  has,  in  the  main,  been  due  to  changes  in  the 
general  conditions  of  life  in  the  different  parts  of  the  United  Kingdom, 
apart  from  the  spread  of  the  practice  of  vaccination,— amongst  other 
things,  to  improvement  of  sanitary  conditions. 

It  is  beyond  doubt  that  an  infectious  disease  like  small-pox  is,  other 
things  being  equal,  more  likely  to  spread  in  towns  than  in  country  districts, 
and  more  likely  to  spread  in  crowded  town  districts  than  in  others  not  so 
densely  populated  ;  so  that  we  should  expect  a  lessened  proportion  of 
overcrowded  dwellings,  by  diminishing  the  opportunities  for  contagion, 
to  check  the  prevalence  of  the  disease  and  consequently  to  render  its 
mortality  less. 

Effect    of   Isolation. 

It  has  been  maintained  that  the  decline  in  small-pox  mortality  is 
largely  due  to  more  frequent  and  systematic  attempts  to  isolate  those 
suffering  from  small-pox.  We  think  an  answer  to  this  contention  is  to  be 
found  in  the  fact  that  it  is  only  in  quite  recent  years  that  there  has  been 
any  systematic  practice  of  isolating  small-pox  patients,  and  that  it  has 
been  confined  even  then  to  a  very  limited  number  of  localities.  The 
fact  to  which  we  are  about  to  call  attention  in  greater  detail  than 
hitherto,  that  the  decline  in  the  deaths  from  small-pox  is  -found  almost 
exclusively  among  those  of  tender  years,  appears  also  to  militate  against 
the  contention.  The  risk  of  contagion  is  not  confined  to  children. 
Adults  also  are  subject  to  it.  If  a  better  system  of  isolation  had  been 
a  main  cause  of  the  reduced  mortality,  we  should  have  expected  to  see  it 
operate  in  the  case  of  adults  as  well  as  of  children.  At  the  same  time 
we  are  far  from  thinking,  as  will  appear  when  we  come  to  deal  with 
that  subject,  that  the  efforts  at  isolation  which  have  characterised  recent 
years  have  been  without  a  beneficial  effect  on  small-pox  mortality. 

Sanitary    Legislation. 

We  have  already  pointed  out  that  on  a  priori  grounds  it  is  reasonable 
to  think  that  improved  sanitary  conditions  would  tend  to  diminish 
the  fatality  of,  and  so  to  a  corresponding  extent  the  mortality  from, 
small-pox.  And  there  can  be  no  doubt  that  the  period  with  which 
we  are  dealing  has  been  characterised  by  an  improvement  of  this 
description.  There  has  been  better  drainage,  a  supply  of  purer  water, 
and  in  other  respects  more  wholesome  conditions  have  prevailed. 

It  may  be  useful  at  this  point  to  furnish  a  brief  summary  of  the 
principal  Sanitary  Acts  which  have  been  passed  relating  to  the  different 
parts  of  the  United  Kingdom. 

In  1848  was  passed  the  first  great  and  comprehensive  measure  which 
may  be  called  the  groundwork  of  our  sanitary  legislation  as  regards 
England.  The  Public  Health  Act  of  1848  was,  however,  principally 
designed  for  towns  and  populous  places  in  England  and  Wales,  not 
including  the  Metropolis,  which  was  dealt  with  in  Acts  passed  in  the 
same  year.  The  powers  of  local  government  supplied  by  the  Act  were 
generally  an  extension  of  those  before  given  by  sundry  local  Acts  to 


REPORT    OF  THE   ROYAL   VACCINATION   COMMISSION.          677 

-Commissioners  of  Sewers  in  the  Metropolis,  and  to  authorities  in  a  few 
large  towns.  Many  provisions  corresponding  to  sections  in  the  Towns 
Improvement  Clauses  Act  of  1847  are  found  in  the  Public  Health  Act, 
and  communities  were  thus  enabled  to  obtain  by  a  simple  process  powers 
which  they,  could  not  previously  obtain  except  by  a  local  Act  incorporating 
sections  of  the  Towns  Improvement  Clauses  Act. 

In  1848  was  also  passed  the  Nuisances  Removal  and  Diseases  Preven- 
tion Act  of  that  year,  in  substitution  for  a  similar  Act  of  1846  which  was 
about  to  expire  ;  and  in  1849  this  Act  of  1848  was  amended.  The 
provisions  of  all  these  three  Acts  extended  to  England,  Scotland  and 
Ireland.  In  1855  a  comprehensive  Nuisance  Removal  Act  was,  as  regards 
England,  substituted  for  the  Acts  passed  in  1848  and  1849 ;  and  in  the 
following  year  there  was  similar  legislation  for  Scotland.  In  1860  the 
English  Act  was  amended  ;  and  in  1866,  by  the  Sanitary  Act  of  that  year 
(to  which  we  shall  again  refer),  the  provisions  of  the  English  Acts  of  1855 
and  1860,  as  then  amended,  were  applied  to  Ireland. 

In  1855,  by  the  Metropolitan  Local  Management  Act  of  that  year, 
provision  was  made  for  the  appointment  of  a  medical  officer  of  health 
and  an  inspector  of  nuisances  by  every  vestry  and  district  board  in 
the  Metropolis.  This  provision  did  not  extend  to  the  City  of  London, 
where,  in  1848,  a  medical  officer  of  health  had  been  appointed  under 
power  given  by  a  local  Act. 

In  1858,  the  Local  Government  Act  of  that  year,  to  be  construed  with 
the  Public  Health  Act  of  1848  as  one  Act,  was  passed,  and  took  effect  in 
all  places  where  that  Act  was  in  force  at  the  time  of  its  passing  ;  and, 
as  regards  England,  these  two  Acts  together  constituted  until  1872  the 
principal  sanitary  legislation  on  the  statute-book. 

There  followed,  however,  within  the  next  ten  years  many  public  Acts 
having  sanitary  objects,  some  applying  to  all,  and  some  to  particular,  parts 
of  the  United  Kingdom,  besides  numerous  other  Acts  of  local  application. 
We  need  only  now  specially  refer  to  one  of  these  public  statutes — the 
Sanitary  Act  of  1866,  which  was  probably  the  most  important,  and  applied, 
in  part  at  least,  to  England,  Scotland  and  Ireland.  This  Act,  amongst 
other  things,  extended  the  powers  of  local  authorities  for  the  disposal  of 
sewage,  and,  in  amending  the  English  Nuisances  Removal  Acts  of  1855 
and  1860,  added  to  the  definitions  of  nuisances,  especially  as  regards 
crowded  houses  and  workshops,  and  to  the  duties  and  powers  of  local 
authorities  for  their  abatement,  especially  in  the  way  of  providing  means 
for  disinfection  and  places  for  the  reception  of  dead  bodies. 

In  1867  the  Public  Health  (Scotland)  Act  was  passed — a  comprehensive 
measure  which  consolidated  into  one  Act,  with  certain  amendments,  the 
whole  statute  law  relating  to  the  public  health  in  Scotland. 

In  1872  a  complete  distribution  of  England  into  sanitary  districts  took 
place,  and  some  further  amendments  were  made  in  the  sanitary  laws.  In 
1875  these  laws  were  consolidated  in  the  Act  of  that  year.  In  1891 
a  Sanitary  Act  was  passed  relating  to  the  Metropolis. 

In  1874  an  Act  was  passed  for  Ireland,  containing  substantially  the 
same  provisions  as  those  which  had  been  enacted  in  the  case  of  England 
in  1872. 


678  SUPPLEMENTARY  APPENDIX. 

Value  of  Vaccination. 

We  have  not  disregarded  the  arguments  adduced  for  the  purpose  of 
showing  that  a  belief  in  vaccination  is  unsupported  by  a  just  view  of  the 
facts.  We  have  endeavoured  to  give  full  weight  to  them.  Having  done  so, 
it  has  appeared  to  us  impossible  to  resist  the  conclusion  that  vaccination 
has  a  protective  effect  in  relation  to  small-pox. 

We  think  :— 

1.  That  it  diminishes  the  liability  to  be  attacked  by  the  disease. 

2.  That  it  modifies  the  character  of  the  disease,  and  renders  it 

(«)  less  fatal,  and  (6)  of  a  milder  or  less  severe  type. 

3.  That  the  protection  it  affords  against  attacks  of  the  disease  is 

greatest  during  the  years  immediately  succeeding  the  operation 
of  vaccination.  It  is  impossible  to  fix  with  precision  the 
length  of  this  period  of  highest  protection.  Though  not  in 
all  cases  the  same,  if  a  period  is  to  be  fixed,  it  might,  we  think, 
fairly  be  said  to  cover  in  general  a  period  of  nine  or  ten  years. 

4.  That  after  the  lapse  of  the  period  of  highest  protective  potency, 

the  efficacy  of  vaccination  to  protect  against  attack  rapidly 
diminishes,  but  that  it  is  still  considerable  in  the  next  quin- 
quennium, and  possibly  never  altogether  ceases. 

5.  That  its  power  to  modify  the  character  of  the  disease  is  also 

greatest  in  the  period  in  which  its  power  to  protect  from 
attacks  is  greatest ;  but  that  its  power  thus  to  modify  the 
disease  does  not  diminish  as  rapidly  as  its  protective  influence 
against  attacks,  and  its  efficacy  during  the  later  periods  of  life 
to  modify  the  disease  is  still  very  considerable. 

6.  That  re -vaccination  restores  the  protection  which  lapse  of  time 

has  diminished;  but  the  evidence  shows  that  this  protection 
again  diminishes,  and  that,  to  ensure  the  highest  degree  of 
protection  which  vaccination  can  give,  the  operation  should  be 
at  intervals  repeated. 

7.  That  the  beneficial  effects  of  vaccination  are  most  experienced 

by  those  in  whose  case  it  has  been  most  thorough.  We  think 
it  may  fairly  be  concluded  that  where  the  vaccine  matter  is 
inserted  in  three  or  four  places,  it  is  more  effectual  than 
when  introduced  into  one  or  two  places  only — and  that  if  the 
vaccination  marks  are  of  an  area  of  half  a  square  inch,  they 
indicate  a  better  state  of  protection  than  if  their  area  be  at  all 
considerably  below  this. 

Question  of  Specific  Protection  or  of  Antagonism. 
When  an  attack  of  disease  secures  immunity  or  protection  against 
another  attack  of  disease,  the  two  attacks  are,  as  a  rule,  attacks  of  the 
same  disease.  Some  pathologists  have,  it  is  true,  of  late  years  been  led  to 
suppose  that  one  disease  may  confer  some  degree  of  immunity  or  protec- 
tion against  another  different  disease  ;  but  instances  of  this  are  few,  andr 
moreover,  cannot  be  regarded  as  thoroughly  established.  The  ordinary 
instances  of  immunity  are  so  clearly  those  in  which  the  attack,  natural  or 


REPORT   OF  THE   ROYAL   VACCINATION   COMMISSION.          679 

artificial,  which  confers  the  immunity  is  of  the  same  disease  as  that  towards 
which  immunity  is  conferred,  that  identity  of  disease  has  been  considered 
as  essential  to  the  conferring  of  immunity.  And  it  has  been  argued  that 
it  is  a  priori  improbable  that  cow-pox  should  confer  immunity  from  small- 
pox, seeing  that  the  two  are  different  diseases.  Such  a  purely  theoretical 
argument  can  have  little  weight  against  positive  evidence  of  vaccination 
having  actually  conferred  immunity.  If  this  be  definitely  proved  to  be  the 
fact,  proof  is  thereby  at  the  same  time  afforded  that  the  theory  is  unsound, 
either  because  a  particular  disease  may  confer  immunity  against  a  different 
disease,  or  because  small-pox  and  cow-pox  are  not  different  diseases.  For 
the  practical  object  with  which  alone  we  are  concerned,  it  is  not  material 
that  we  should  reach  any  conclusion  upon  the  question  what  is  the  real 
source  of  error  in  the  theory  alluded  to,  supposing  it  to  be  erroneous  ? 
We  shall  content  ourselves,  therefore,  with  a  very  brief  notice  of  the 
subject. 

It  appears  to  us  that  we  may  dismiss  for  practical  purposes  the 
theoretical  questions  which  were  discussed  before  us  so  fully.  If  the  fact 
be  established  that  the  introduction  of  vaccine  matter  and  the  consequent 
vaccinia  produce  some  effect  upon  the  human  body  which  renders  it  less 
susceptible  to  small-pox,  or  which  modifies  that  disease  when  the  small-pox 
virus  enters  the  system,  it  will  not  be  a  strange  or  unwonted  experience 
that  we  should  be  unable  to  explain  how  this  comes  about.  Science 
has  not  yet  succeeded  in  freeing  therapeutics  or  kindred  subjects  from 
obscurity,  or  in  solving  all  the  problems  which  they  present.  The  precise 
modus  operand},  by  which  a  previous  attack  of  a  disease  furnishes  security 
against  a  subsequent  attack  by  the  same  disease  has  not  yet  been  elucidated. 
There  can  be  no  cause  for  astonishment,  then,  if  we  are  unable  to  trace 
the  steps  by  which  vaccination  exerts  a  protective  influence,  supposing 
the  fact  that  it  does  so  be  established,  nor  is  it  essential  that  we  should 
succeed  in  tracing  them.  Our  inability  to  accomplish  this  does  not  seem 
to  us  to  be  the  slightest  reason  for  regarding  with  doubt  the  conclusions 
to  which  the  facts  lead  us. 

Professor  Crookshank,  than  whom  no  one  has  more  strongly  insisted  on 
the  theoretical  arguments  against  the  protective  influence  of  vaccination  in 
relation  to  small-pox,  gives  it  as  his  opinion  that  vaccination  creates  a 
transient  antagonism  to  that  disease.  We  understand  his  view  to  be  that 
an  attack  of  disease  can  only  afford  protection  against  the  same  disease, 
and  that  small-pox  and  cow-pox  are  not  the  same  but  different  diseases. 
We  gather,  however,  that,  in  his  opinion,  so  long  as  the  state  of  antagonism 
lasts,  the  person  in  whose  system  it  exists  is  less  likely  to  suffer  from 
small-pox  than  he  would  be  if  the  state  of  antagonism  were  wanting. 
This  seems  to  us  to  amount  in  effect  to  the  same  thing  as  saying  that 
during  that  period  vaccination  has  conferred  some  protection.  Whether 
the  effect  be  to  create  antagonism  or  to  confer  protection,  and  whatever 
difference  there  be  between  the  mod  tin  ojtertntdi  in  the  one  case  and  in  the 
other,  we  know  equally  little  about  it.  If  a  condition  of  transient 
antagonism  to  small-pox  is  induced  by  vaccination,  theoretical  considera- 
tions will  not  afford  a  guide  of  the  slightest  value  to  the  conclusions  how 
long  this  transient  antagonism  will  last,  or  how  soon  it  will  pass  away. 


SUPPLEMENTARY   APPENDIX. 

Experience,  and  experience  alone,  can  answer  that  question.  A  priori  we 
-do  not  see  that  there  is  any  better  reason  for  supposing  that  it  would  last 
for  two  or  three  years  than  that  its  duration  would  extend  to  ten  years. 

Cow-pox  and  Small-pox  not  Convertible. 

Jenner  himself,  in  his  first  paper,  advanced  the  view  that  the  cow-pox 
and  small-pox  were  identical  with  each  other ;  and  since  his  time 
numerous  observers  have  attempted  to  prove  the  identity  of  the  two 
diseases  experimentally — namely,  by  giving  rise  to  cow-pox  in  the  cow 
through  the  inoculation  of  small-pox  matter,  or  by  the  introduction  of 
contagion  in  other  ways.  It  may  at  once  be  stated  that  while  cow-pox 
is  readily  transferred  from  the  cow  to  man  and  back  again  from  man  to 
the  cow,  the  disease  in  man  being  identical  with  that  in  the  cow,  small- 
pox cannot  be  transferred  from  man  to  the  cow  so  as  to  give  rise  to  a 
disease  in  the  latter  identical  in  its  features  with  the  small-pox  of  man. 
^Nor  can  cow-pox  be  so  transferred  to  man  as  to  give  rise  in  him  to 
small-pox.  The  two  diseases  are  not  in  this  sense  convertible. 

Small-pox  Vaccine. 

In  most  cases  the  attempt  to  transfer  small-pox  from  man  to  the 
•cow  has  had  simply  a  negative  result  ;  no  obvious  effect  of  any  kind  has 
been  observed.  This  has  been  the  case  in  the  attempts  to  introduce  the 
contagion  through  absorption  by  the  respiratory  or  digestive  organs,  and 
in  most  of  the  attempts  to  introduce  the  contagion  by  inoculation.  In 
certain  instances  these  latter  attempts  have  produced  results  which  may 
be  briefly  described  in  three  categories.  (We  may  pass  over  the  isolated 
experience  of  Thiele,  who  in  1838  asserted  that  by  keeping  small-pox 
matter  sealed  between  glass  plates  for  ten  days  before  using  it,  and 
by  diluting  it  with  milk  when  using  it  for  inoculation,  the  matter  thus 
treated  through  ten  removes  through  the  human  body — the  cow  not 
intervening  at  all — was  converted  into  something  which  gave  results 
identical  with  those  of  ordinary  vaccine  matter.  We  are  not  aware  of 
any  attempt  to  corroborate  this  experiment.) 

The  first  category  includes  the  experiments  in  which  the  inoculation 
of  small-pox  matter  into  the  udder,  or  adjoining  parts,  of  the  cow  gave 
rise  at  or  near  the  seat  of  inoculation  to  a  vesicle,  either  identical  in 
visible  characters  with  the  ordinary  vaccine  vesicle  produced  by  inocula- 
tion with  the  matter  of  cow-pox,  or  to  a  vesicle  the  features  of  which, 
while  not  corresponding  wholly  with  those  of  a  perfect  vaccine  vesicle, 
so  closely  resembled  them  as  to  justify  the  vesicle  being  called  a  vaccine 
vesicle.  Further,  the  matter  from  a  vesicle  which  at  the  first  inoculation 
had  not  the  characters  of  a  perfect  vaccine  vesicle,  when  carried  through 
a  second  or  third  remove  in  the  cow,  fully  acquired  those  characters,  and 
when  transferred  to  man  gave  results  indistinguishable  from  the  ordinary 
vaccine  vesicle.  Indeed,  lymph  of  such  an  origin  has  come  into  general 
use  for  vaccination  purposes.  Of  the  experiments,  the  best  known  or 
most  quoted  are  those  of  Thiele  (1838),  Ceely  (1840),  Badcock  (between 
1840  and  1860),  Voigt  (1881),  Haccius  and  Eternod  (1890),  King  (1891), 


REPORT    OF   THE   ROYAL   VACCINATION    COMMISSION.  681 

Simpson  (1892),  and  Hinie  (1892)  ;  but  there  are  several  others.  The 
details  of  the  experiment  are  very  scanty  in  the  cases  of  Thiele  and 
Badcock,  but  more  full  in  the  others,  especially,  perhaps,  those  of  Ceely 
and  Haccius. 

In  the  second  category  may  be  placed  the  experiments  of  Klein  and 
Copeman.  Klein,  who  had  in  1879  obtained  in  31  trials  what  then 
appeared  simply  negative  results,  found  in  a  renewed  research  in  1892 
that  the  result  of  the  first  inoculation  in  the  cow  of  small-pox  matter 
was  not  a  distinct  vesicle  but  merely  a  thickening  and  redness  of  the 
wound.  Lymph  pressed  from  the  thickened  wound  produced,  when 
inoculated  into  a  second  cow,  a  like  result,  but  rather  more  marked  ; 
the  thickening  and  reddening  still  further  increased  with  a  third  and 
a  fourth  cow.  Lymph  squeezed  from  the  wounds  of  the  fourth  cow 
produced  in  a  child  typical  vaccine,  and  crusts  from  the  child  produced 
typical  vaccine  in  a  cow.  Copeman  obtained  somewhat  similar  results  ; 
the  appearances  increasing  in  three  removes  and  approaching  those  of 
typical  vaccine,  but  not  reaching  them. 

The  third  category  consists  of  -ttye  results  obtained  in  an  elaborate 
inquiry  conducted  by  a  Commission  of  the  Society  of  Medical  Sciences  at 
Lyons,  with  Chauveau  at  its  head.  Those  results,  reported  in  1 865 ,  were 
briefly  as  follows  : — 

Inoculation  of  the  cow  with  small-pox  matter  in  any  one  of  the  30 
animals  used  did  not  give  rise  to  a  vaccine  vesicle.  Nevertheless  a 
definite  result  was  obtained  ;  in  the  form,  however,  not  of  a  vesicle,  but 
of  a  thickening  and  inflammation  of  the  wound  ;  when  a  puncture  was 
employed  this  became  a  papule.  Lymph  squeezed  from  such  a  papule 
and  inserted  into  a  second  animal  gave  rise  to  a  like  papule  :  and  this, 
again,  might  be  used  for  a  third  animal,  but  often  failed  ;  and  the  effect 
could  in  no  case  be  carried  on  through  more  than  three  or  four  removes. 

When  the  inoculation  was  repeated  on  an  animal  in  which  a  previous 
inoculation  had  produced  such  a  papule,  no  distinct  papule  was  formed  ; 
and,  moreover,  lymph  squeezed  from  the  seat  of  inoculation  produced  no 
effect  at  all  when  used  for  the  subsequent  inoculation  of  another  animal. 
This  shows  that  the  development  of  the  papule  was  the  result  of  the 
specific  action  of  the  virus.  The  same  is  shown  by  the  fact  that  no  such 
papule  was  produced  when  the  small- pox  matter  was  inserted  into  an 
animal  which  had  previously  had  cow-pox  naturally  or  artificially,  as  well 
as  by  the  fact  that  when  an  attempt  was  made  to  vaccinate,  with  vaccine 
matter  of  proved  efficacy,  an  animal  on  which  a  papule  had  been  so 
developed  by  inoculation  with  small-pox,  the  vaccination  failed,  though 
the  animal  had  never  had  natural  cow-pox  or  had  never  been  vaccinated. 
The  specific  nature  of  the  lymph  of  the  papule  is  further  shown  by  the 
fact  that  such  lymph  when  used  on  the  human  subject  gave  rise  to  veri- 
table small-pox.  It  has  been  urged  that  in  this  case  the  virus  producing 
the  effect  was  simply  the  old  virus  used  in  the  inoculation,  producing 
the  papule  and  still  clinging  to  the  wound.  This  is  disproved  by  the 
experience  that  lymph  from  a  ))<if>ul<'  <>f  the  second  re  more  also  gave  rise  in 
the  human  subject  to  veritable  small-pox. 

Thus  Chauveau  and  his  Commission  found  that  small-pox  implanted 


682  SUPPLEMENTARY  APPENDIX. 

in  the  cow  gave  rise  to  a  specific  effect  which  was  not  cow-pox  but  was 
of  the  nature  of  small-pox,  though  its  manifestations  in  the  cow  were 
different  from  those  of  small-pox  in  man.  They  also  obtained  similar 
results  in  attempting  to  transfer  small-pox  to  the  horse. 

It  must  be  admitted  that  the  results  finally  obtained  in  some  of  the 
successful  cases  were  indistinguishable  from  those  of  vaccination  ;  the 
characters  of  the  local  vesicle,  the  absence  of  eruptive  pustules  and  of 
contagiousness,  show  that  the  lymph  thus  apparently  originating  from 
small-pox  in  the  hands  of  Ceely,  Badcock,  and  others,  was  vaccine  lymph. 
It  has  been  urged  that  a  vaccine  vesicle  making  its  appearance  in  the 
wound  of  inoculation  with  small-pox  was  due  to  the  accidental  intro- 
duction of  cow-pox  matter  into  the  wound  ;  the  small-pox  matter  in 
the  wound  produced  no  effect,  and  the  cow-pox  matter  its  usual  effect. 
Several  considerations  support  this  view.  The  cow  is  peculiarly  suscep- 
tible to  cow-pox.  In  some  cases  (Ceely,  Voigt),  the  animal  was 
vaccinated  as  well  as  inoculated  with  small-pox  :  thus,  in  Ceely's  first 
case,  the  animal  was  inoculated  with  small-pox  on  one  side  of  the  body, 
and  a  few  days  after  vaccinated  on  the  other  side.  In  many  cases  the 
experiments  were  conducted  in  an  animal  vaccine  establishment,  the 
stalls,  the  operating  tables,  and  the  assistants  being  those  used  or  engaged 
in  vaccination.  It  is  true  that  in  some  cases  at  least  special  precautions, 
sterilisation  of  instruments  and  the  like,  were  taken  to  avoid  the  accidental 
introduction  of  cow-pox  ;  but  in  observations  of  this  kind  the  difficulties 
of  avoiding  all  such  sources  of  error  are  notorious.  Still  the  successful 
cases  are  now  so  numerous  that  it  is  difficult  to  resist  the  conclusion  that 
the  same  accident  could  not  have  occurred  in  all,  and  that  a  transformation 
of  small-pox  into  cow-pox — that  is  to  say,  into  the  artificially  inoculated 
cow-pox  which  we  call  vaccine  * — really  took  place. 

Accepting  this  view  provisionally,  it  may  be  remarked  that  in  most 
cases  the  transformation  was  sudden  and  complete  ;  the  small-pox  virus, 
under  the  influence  of  the  tissues  of  the  cow,  became  immediately 
converted  into  vaccine  virus,  and  this  produced  a  typical  vaccine  vesicle. 
In  some  cases  (ex.  gr.,  that  of  Hime)  the  transformed  virus  produced  its 
effect  not  in  the  wound  of  inoculation,  or  not  chiefly  so,  but  at  some  little 
distance  from  it.  In  some  cases  the  characters  of  the  vesicle  first  formed, 
though  sufficiently  distinct  to  justify  the  vesicle  being  called  a  vaccine 
vesicle,  were  not  those  of  a  perfect  vaccine  vesicle,  but  the  lymph  from 
such  a  vesicle,  at  least  after  one  or  two  removes,  gave  rise  to  most  typical 
vaccine  vesicles. 

In  Klein's  experiments  the  transformation  was  gradual.  In  his  fourth 
cow,  the  virus  was  as  yet  not  typical  vaccine,  since  it  did  not  produce  a 
typical  vesicle  ;  yet  it  was  so  far  already  vaccine  that,  transferred  to  the 
child,  it  produced  typical  vaccine  (unless  we  suppose  some  accidental 
introduction  of  vaccine  to  have  taken  place).  That  the  vesicle  on  the 
child  was  vaccine,  and  not  small-pox  unaccompanied  by  eruptive  pustules, 
was  shown  not  only  by  its  characters  but  also  by  the  fact  that  lymph  from 
it  produced  typical  vaccine  in  the  cow. 

In  "Chauveau's  experiments  no  transformation  at  all  took  place. 
*  The  italics  are  mine. — E.M.C. 


REPORT    OF   THE   ROYAL   VACCINATION   COMMISSION.          683 

As  has  been  urged  in  another  place,  there  are  no  adequate  reasons- 
leading  us  to  believe  that  in  the  human  subject  the  small-pox  virus  and 
the  cow-pox  virus  can  so  act  on  each  other  as  to  form  a  hybrid  disease. 
But  this  does  not  preclude  the  view  that,  accepting  the  conclusion  that 
the  body  of  the  cow  has  the  power  to  convert  small-pox  into  vaccine,  the 
virus  may  exist  for  a  while  in  a  phase  in  which,  while'  ceasing  to  be 
typical  small-pox,  it  has  not  yet  fully  acquired  the  characters  of  vaccine, 
and  we  may  regard  Klein's  results  as  illustrating  this.  In  some  of  the 
experiments — for  instance,  those  of  Ceely  and  Voigt — the  results  obtained 
with  the  lymph  of  the  vesicle  produced  by  the  inoculation  of  small-pox 
give  rise  to  the  suspicion  that  the  lymph  had  small-pox  qualities,  as  seenr 
for  example,  in  the  case  of  Ceely's  assistant,  Taylor  ;  but  the  facts  cannot 
be  said  to  be  more  than  suspicious — they  are  not  decisive.  Moreover, 
admitting  that  the  vesicle  itself  in  such  cases  was  the  result  of  the  trans- 
formed virus,  some  not  transformed  old  virus  might  still  remain  dormant 
in  the  wound,  and  might  be  present  in  the  lymph  of  the  vesicle,  mixed 
with  the  transformed  and  generating  virus  ;  this  old  virus  might  have 
happened  to  be  in  excess  on  the  point  of  the  lancet  which  wounded 
Taylor. 

Small-pox   Vaccine — Cow-pox   Vaccine — Horsebox   Vaccine — Cattle- 
plague  Vaccine — Sheep-pox  Vaccine. 

Taking  all  the  various  facts  into  consideration,  we  seem  led  to  the 
provisional  conclusion  that  under  certain  conditions  the  tissues  of  the 
cow  are  able  to  transform  small-pox  into  vaccine,  that  these  conditions 
may  be  such  as  to  lead  to  the  transformation  being  sudden  and  complete, 
that  under  certain  other  conditions  the  transformation  may  be  gradual 
and  incomplete,  and  that  under  certain  other  conditions  (and  these  seem 
most  commonly  to  obtain)  the  transformation  into  vaccine  does  not  take 
place  at  all.  But  what  the  above  conditions  are  has  not  as  yet  been 
clearly  made  out.  It  has  been  suggested  that  one  condition  favourable 
to  the  transformation  is  extreme  youth  of  the  subject :  to  effect  the 
change  the  animal  used  should  be  a  calf  of  not  more  than  three  or  four 
months  old  :  but  this  is  not  definitely  proved. 

Until  these  favourable  conditions  have  been  clearly  recognised,  so  that, 
the  conditions  being  fulfilled,  the  transformation  will  always  be  secured, 
the  conclusion  cannot  be  regarded  as  indisputable.  Moreover,  it  must 
be  borne  in  mind  that  effects  more  or  less  closely  resembling  a  vaccine 
vesicle  have  been  at  various  times  obtained  by  various  observers  through 
inoculating  man  or  the  cow  or  another  animal  with  material  other 
than  that  obtained  from  the  pustules  of  the  small-pox  of  man.  Much 
discussion  has  taken  place  concerning  the  "  grease  "  of  the  horse,  which 
Jenner  believed  to  be  the  origin  of  the  cow-pox  of  the  cow.  Without 
entering  into  any  discussion  of  the  matter,  it  may  be  said  that  investiga- 
tion has  shown  that  horses  do  suffer  from  a  malady  which,  transferred  to 
the  cow,  gives  rise  to  a  vaccine  identical  apparently  with  that  produced 
by  the  inoculation  of  the  natural  cow-pox.  Hence  this  malady  is  spoken 
of  as  the  "  horse-pox,"  and  some  cases  at  least  of  so-called  "  grease  " 
appear  to  be  cases  of  this  horse-pox.  But  it  is  at  least  not  proved  that 


684  SUPPLEMENTARY  APPENDIX. 

all  the  cases  of  "  grease  "  which  by  inoculation  were  found  to  give  rise  to 
vaccine  vesicles  in  man  were  cases  of  true  horse-pox.  And  this  at  least 
must  be  said,  that  no  investigations  as  complete  and  varied  as  those  which 
have  been  carried  out  with  regard  to  the  development  of  vaccine  vesicles 
through  the  inoculation  of  small-pox  matter,  have  been  carried  out  with 
regard  to  the  alleged  development  of  vaccine  vesicles  by  tho  inoculation 
of  other  material,  such  as  the  matter  from  the  eruptions  of  the  sheep-pox, 
the  cattle  plague,  and  the  like.  Nor  have  there  been  like  extended  in- 
quiries as  to  the  production  of  vesicles  resembling  those  of  vaccine  by  the 
inoculation  of  small-pox  matter  into  animals  other  than  the  cow  or  the 
horse  ;  such  results  as  have  been  obtained  by  observers  are  conflicting. 
'There  is  still  room  for  much  inquiry  ;  meanwhile  it  may  be  said  that,  in 
any  case,  the  evidence  in  favour  of  a  possible  transformation  of  small-pox 
into  vaccine  is  sufficiently  strong  to  remove  the  force  of  the  theoretical 
objection  to  the  power  of  vaccination  to  secure  immunity  towards  small- 
pox, on  the  ground  that  the  two  diseases  are  absolutely  distinct. 

/ 
Risks  of  Vaccination. 

It  must  not  be  forgotten  that  the  introduction  into  the  system  of  even 
a  mild  virus,  however  carefully  performed,  is  necessarily  attended  by  the 
production  of  local  inflammation  and  of  febrile  illness.  If  these  results 
did  not  in  some  measure  follow,  the  practice  would  probably  fail  in  its 
protective  influence.  As  a  rule,  the  inflammation  and  illness  are  of  a 
trifling  character  ;  in  exceptional  cases,  however,  they  may  exhibit  more 
severity,  and,  as  certain  facts  submitted  to  us  in  evidence  have  shown, 
there  are  cases,  though  these  are  rare,  where  a  general  eruption  may 
follow  vaccination. 

In  order  to  determine  how  far  the  risk  of  erysipelas  is  a  necessary 
incident  of  vaccination,  what  is  the  extent  of  that  risk,  and  how  it  may 
best  be  avoided,  it  is  necessary  to  consider  the  various  circumstances 
which  may  occasion  erysipelas  and  allied  diseases  in  the  case  of  vaccinated 
children.  It  is  established  that  lymph  contains  organisms,  and  may 
contain  those  which  under  certain  circumstances  would  be  productive  of 
erysipelas.  It  is  therefore  possible  that  some  contagious  material  (the 
specific  virus  of  erysipelas,  for  instance,)  may  be  conveyed  at  the  time  of 
vaccination,  owing  either  to  its  presence  in  the  lymph  employed,  or  to  its 
being  conveyed  by  the  vaccinator  himself,  or  by  those  with  whom  the 
child  comes  in  contact  at  ,the  time  of  vaccination.  We  believe  that  the 
cases  in  which  the  virus  is  conveyed  at  the  time  of  vaccination  are  rare. 
It  has,  however,  in  some  instances  been  clearly  established,  the  immediate 
occurrence  of  erysipelas  in  several  co-vaccinees  making  it  practically 
certain  that  some  virus  was  conveyed  at  the  time  of  the  operation.  In 
some  instances  where  this  has  been  the  case,  and  there  is  every  reason  for 
believing  that  the  contagion  was  conveyed  through  the  medium  of  the 
lymph,  it  is  nevertheless  in  evidence  that  the  vaccinifer  did  not  display 
anything  more  than  a  slightly  inflamed  arm.  The  scrupulous  avoidance 
of  inflamed  arms  in  vaccinifers  will  do  much  to  reduce  the  risk  of 
•conveying  erysipelas  in  the  act  of  vaccination  (a  risk  which,  as  we  have 


REPORT   OF   THE   ROYAL   VACCINATION   COMMISSION.  685 

seen,  has  been  proved  to  be  a  very  slight  one),  but  it  is  possible  it  would 
not  wholly  remove  it. 

We  have  dwelt  upon  features  presented  by  the  cases  of  erysipelas  and 
various  forms  of  septic  disease  which  have  followed  vaccination,  because 
they  suggest  precautions  which  may  be  adopted  to  lessen,  if  not  to 
prevent,  such  evils  in  the  future.  If,  for  example,  vaccination  were 
performed  at  the  patient's  home  instead  of  at  a  public  vaccination  place, 
the  chance  of  disease  being  contracted  at  the  time  of  vaccination  would 
be  to  some  extent  diminished  ;  and  the  same  may  be  said  of  the  inspection 
of  the  vaccinated  person  which  takes  place  eight  days  after  the  operation. 
On  these  points  we  shall  have  some  remarks  and  recommendations  to 
make  at  a  later  stage  of  our  report. 

A  study  of  the  cases  which  have  been  made  the  subject  of  careful 
examination  and  report  points  to  the  conclusion  that  an  exercise  of 
greater  care  would  largely  diminish  the  risk,  already  small,  of  erysipelas- 
contagion  and  blood-poisoning. 

Although  it  may  be  confidently  hoped  that  by  additional  care  on  the 
part  both  of  vaccinators  and  parents,  the  number  of  inflamed  arms  and 
of  cases  of  erysipelas  may  be  reduced  to  very  few,  yet  it  is  not  to  be 
expected  that  such  occurrences  will  be  wholly  prevented.  A  vaccination 
wound  is,  like  one  from  anv  other  cause,  so  long  as  it  exists,  a  source  of 
some  risk. 

The  use  of  calf-lymph,  though  it  may  be  supposed  to  be  more  free 
from  the  risk  of  conveying  erysipelas,  does  not  appear  to  prevent  inflamed 
arms.  Some  witnesses  have  indeed  supposed  that  it  is  attended  with 
more  risk  of  inflammation  than  the  employment  of  that  taken  from  the 
human  subject.  This  opinion  has  not,  however,  been  corroborated  by 
some  of  those  of  widest  experience. 

Nothing  has  produced  so  deep  an  impression  hostile  to  vaccination  as 
the  apprehension  that  syphilis  may  be  communicated  by  it.  It  was  at 
one  time  doubted  whether  syphilis  could  result,  and  it  was  even  confi- 
dently asserted  that  it  could  not.  The  fact  that  this  was  possible  had 
been  fully  established,  and  was  generally  acknowledged  by  the  medical 
profession,  before  we  commenced  our  inquiries. 

The  very  close  resemblance  in  certain  very  rare  cases  of  the  results 
of  vaccination,  whether  with  calf-lymph  or  humanised  lymph,  to  those 
attributed  to  syphilis  (a  resemblance  so  close  that  it  has  caused  in  a  few 
cases  a  difference  of  opinion  whether  the  disease  was  syphilis  or  vaccinia) 
has  led  to  the  expression  by  Dr.  Creighton  of  the  opinion  that  there  is 
some  essential  relationship  between  the  two  diseases.  This,  however, 
is  a  point  of  speculative,  almost  it  might  be  said  of  transcendental 
pathology,  upon  which  for  practical  purposes  it  is  useless  to  enter.  It 
must  be  sufficient  to  remark  that,  whatever  may  be  the  relationship 
alluded  to,  it  exists,  if  it  exists  at  all,  equally  between  small-pox  and 
syphilis  as  between  vaccination  and  syphilis.  For  all  practical  purposes 
variola  and  vaccinia  are  both  wholly  distinct  from  syphilis,  and  their 
differences  are,  with  the  rarest  exceptions,  easily  recognised.  They  are 
alike  in  being  attended  by  affections  of  the  skin  and  mucous  membranes, 
and  exceptionally  by  disease  of  the  bones,  eyes,  and  other  parts  ;  but  in  all 


686  SUPPLEMENTARY   APPENDIX. 

these  it  is  a  question  of  resemblance  and  not  of  identity  with  which  we 
have  to  deal. 

Only  a  few  items  of  the  evidence  produced  before  us  appear  to  require 
special  notice  :  among  these,  the  most  prominent  is  what  has  been  known 
as  the  "  Leeds  case,"  upon  which  we  have  heard  the  evidence  of  Mr. 
"Ward,  Mr.  Littlewood  and  Dr.  Barrs.  The  witnesses  named  regarded  it  as 
a  case  of  syphilis,  conveyed  by  vaccination,  but  all  of  them  admitted  that 
the  course  of  events  was  most  unusual.  We  have  carefully  investigated 
this  case,  and  notwithstanding  the  opinion  formed  by  the  witnesses,  there 
appears  good  reason  to  doubt  whether  it  was  one  of  syphilis.  The  case  was 
made  the  subject  of  careful  inquiry  by  Dr.  Barlow  on  our  behalf,  who 
shared  the  doubt  we  have  expressed.  The  view  taken  by  the  medical 
inspector  of  the  Local  Government  Board  who  in  the  first  instance  in- 
vestigated the  case  was  that  it  was  a  case  of  hereditary  syphilis.  It  seems 
certain,  however,  that  the  parents  of  the  child  whose  death  was  in  question 
were  not  in  any  way  affected  with  syphilis.  The  vaccinifer  also  appeared 
to  be  free  from  any  taint  of  that  disease,  and  its  family  history  confirmed 
this  view.  The  co-vaccinees  from  the  same  lymph  also  exhibited  no  trace 
of  syphilis.  These  facts  of  themselves  make  out  a  strong  case  against 
that  having  been  the  nature  of  the  disease.  Coupled  with  the  fact  that 
it  could  not  have  been  communicated  by  the  vaccinator  himself,  they  seem 
to  render  it  practically  impossible  that  syphilis  was  the  cause  of  death. 
If  the  symptoms  exhibited  had  in  all  respects  corresponded  with  those 
which  are  known  to  characterise  syphilis,  the  proper  inference  might  have 
teen  that  there  was  some  error  in  ascertaining  the  facts  of  the  case.  But 
it  is  beyond  question  that  the  course  of  events  was  very  different  in  some 
respects  from  that  experienced  in  undoubted  cases  of  syphilis,  and  we 
think  the  true  conclusion  is  that  it  was  not  a  case  of  that  disease.  It  may 
probably  be  classed  with  a  few  others  as  examples  of  gangrene  and  blood- 
poisoning,  the  direct  result  of  vaccination,  which  are  not  to  be  explained 
by  supposing  the  introduction  of  any  syphilitic  or  other  poison.  Fortu- 
nately, such  cases  are  extremely  rare — so  much  so  that  the  witnesses 
concerned  knew  of  no  case  precisely  parallel. 

The  evidence  offered  to  us  would  lead  to  the  belief  that,  whilst  with 
ordinary  care  the  risk  of  communication  of  syphilis  in  the  practice  of  arm- 
to-arm  vaccination  can  for  the  most  part  be  avoided,  no  degree  of  caution 
can  confer  an  absolute  security.  The  rejection  as  vaccinifers  of  young 
infants,  say  below  four  months  of  age  (in  whom  congenital  syphilis  may 
be  as  yet  undeclared),  and  of  adults  (in  whom  the  disease  may  possibly 
have  been  recently  acquired)  are  precautions  which  would  probably  shut 
out  almost  the  whole  of  the  risk.  The  outbreaks  of  syphilis  in  connection 
with  vaccination  which  have  been  mentioned  to  the  Commission  (all  of 
which  had  been  previously  published)  have  occurred  chiefly  in  arm-to-arm 
vaccination  amongst  soldiers,  or  from  the  use  as  vaccinifers  of  young 
infants  the  offspring  of  parents  whose  history  was  not  known  to  the 
vaccinator.  It  must,  however,  be  admitted  that  neither  the  examination 
of  the  vaccinifer  if  taken  alone,  and  without  a  knowledge  also  of  the 
parents,  nor  the  most  scrupulous  avoidance  of  any  visible  admixture  of 
blood  with  the  lymph,  are  in  themselves,  however  valuable,  sufficient 


REPORT   OF   THE   ROYAL   VACCINATION   COMMISSION.  687 

absolutely  to  exclude  risk.  The  evidence  given  by  Dr.  Husband,  of  the 
Vaccine  Institution  of  Edinburgh,  established  the  fact  that  all  lymph, 
however  pellucid,  does  really  contain  blood  cells.  Absolute  freedom  from 
risk  of  syphilis  can  be  had  only  when  calf-lymph  is  used  ;  though  where 
the  antecedents  of  the  vaccinifer  are  fully  ascertained,  and  due  care  is 
used,  the  risk  may  for  practical  purposes  be  regarded  as  absent. 

It  is  obvious  that  the  employment  of  calf -lymph  only  would  wholly 
exclude  the  risks  as  regards  both  syphilis  and  leprosy.  Respecting  the 
latter  disease,  however,  there  appears  to  be  reason  to  doubt  whether  any 
risk  exists,  and  at  any  rate  it  .does  not  concern  the  British  population. 
Even  in  leprosy  districts  the  employment  of  English  human  lymph  would 
be,  so  far  as  leprosy  is  concerned,  as  safe  as  that  from  the  calf. 

There  can  be  no  doubt  that  vaccination  ought  to  be  postponed  when 
erysipelas,  scarlet  fever,  measles,  or  chicken-pox  are  prevalent  in  the 
neighbourhood  of  the  child's  residence,  or,  if  the  child  is  not  to  be 
vaccinated  at  home,  either  there  or  near  the  place  of  vaccination.  Here 
again  there  would  be  a  gain  if  the  home  was  more  often  the  place  of 
vaccination. 

It  would,  in  our  opinion,  be  an  advantage  if  the  postponement  of 
vaccination  were  expressly  permitted,  not  only  on  account  of  the  state  of 
the  child,  but  of  'its  surroundings  and  any  other  conditions  rendering  the 
operation  at  the  time  undesirable.  If  more  discretion  in  this  respect 
were  possessed  and  exercised,  we  think  untoward  results  would  become 
even  rarer  than  they  are. 

We  are  quite  alive  to  the  objections  which  may  be  urged  against  a 
prolongation  of  the  period  within  which  vaccination  must  be  performed. 
It  will  naturally  be  said  that  a  number  of  children,  who  otherwise  would 
be  protected  against  small-pox,  would  be  left  without  that  protection, 
and  would  thus  be  liable  to  suffer  from  the  disease  themselves,  and  be  a 
source  of  danger  to  others.  It  must  be  remembered,  however,  that  so 
long  as  children  cannot  walk,  the  risk  of  their  contracting  contagion  is 
less  than  if  they  were  able  to  move  freery  about  and  mix  with  other 
people,  and  that,  for  the  same  reason,  the  risk  of  their  communicating 
contagion  to  others  is  less.  We  cannot  trace  in  the  statistics  relating  to 
Scotland  any  grounds  for  believing  that  the  later  compulsory  vaccination 
age  which  prevails  in  that  country  as  compared  with  England  has  affected, 
to  any  substantial  extent,  the  general  small-pox  mortality  of  Scotland, 
though  no  doubt  it  may  have  led  to  some  deaths  among  children  under 
six  months  of  age  which  otherwise  would  not  have  taken  place. 

We  have  already  shown  how  satisfactory  a  position  Germany  has 
occupied  in  relation  to  small-pox  since  the  year  1874.  The  age  of  com- 
pulsion in  that  country  is  the  end  of  the  next  calendar  year  after  birth. 
It  is  true  that  re- vaccination  has  been  there  made  compulsory  as  well  as 
primary  vaccination  ;  but  we  think  the  experience  of  Germany  is  not 
without  its  bearing  on  the  question  we  are  now  considering.  Wherever 
the  line  is  drawn,  whether  at  three  months  or  six  months,  it  will  always 
leave  a  class  of  unvaccinated  persons.  The  age  to  be  fixed  is  a  question 
of  policy  into  which  many  considerations  must  enter.  If  an  extension 
of  the  age  within  which  vaccination  was  required  rendered  its  untoward 


688  SUPPLEMENTARY  APPENDIX. 

incidents  fewer  in  number,  and  diminished  hostility  to  the  operation,  it 
may  be  that  on  the  whole  it  would  promote  the  cause  of  vaccination,  and 
secure,  as  its  result,  that  the  number  of  vaccinated  persons  would  be 
greater  than  at  present. 

Means,  other  than  Vaccination,  for  diminishing  the  Prevalence  of  Small-pox  ,- 
and  how  far  such  means  could  be  relied  on  in  place  of  Vaccination. 

Another  question  upon  which  we  are  asked  to  report  is,  what  means, 
other  than  vaccination,  can  be  used  for  diminishing  the  prevalence  of 
small-pox  ;  and  how  far  such  means  could  be  relied  on  in  place  of 
vaccination. 

The  means,  other  than  the  inoculation  of  small-pox  or  cow-pox,  which 
have  been  referred  to  by  witnesses  as  being  capable  of  diminishing  the 
prevalence  of  small-pox,  are  such  means  as  have  been  employed  against 
infectious  diseases  generally  ;  they  may  be  summarised  as — (1)  Measures 
directed  against  infection,  e.g.,  prompt  notification,  isolation  of  the 
infected,  disinfection,  etc.  ;  (2)  Measures  calculated  to  promote  the  public 
health,  the  prevention  of  overcrowding  in  dwellings  or  on  areas, 
cleanliness,  the  removal  of  definite  insanitary  conditions,  etc. 

The  principle  underlying  the  practice  of  isolation  with  its  accompany- 
ing machinery  is  obviously  the  very  opposite  of  that  which  recommended 
the  practice  of  inoculation  ;  it  aims  at  exclusion  of  the  disease,  whereas 
inoculation  aimed  at  universal  acceptance  by  artificially  "  sowing  "  or 
"  buying "  the  disease.  Except  in  regard  to  the  plague,  our  knowledge 
and  practice  of  measures  of  isolation  and  quarantine  against  epidemics  is 
of  relatively  recent  growth.  As  the  result  of  increased  knowledge  of  the 
mode  of  propagation  of  infectious  diseases,  of  greater  sanitary  activity, 
and  under  the  stimulus  of  legislation,  organised  effort,  more  or  less 
thorough,  is  now,  in  this  as  in  other  countries,  directed  against  the  spread 
of  dangerous  infectious  diseases.  Side  by  side  with  a  vaccination  system,, 
means  of  isolation,  etc.,  have  been  successfully  employed  to  check  the 
spread  of  small-pox. .  They  have  also  been  sometimes  so  employed  in 
recent  years  in  places  where  vaccination  has  fallen  into  disuse. 

It  will  be  well  to  commence  with  a  brief  statement  of  the  growth  of 
our  knowledge  on  the  subject  of  isolation  as  a  means  of  dealing  with 
infectious  or  contagious  diseases.  We  have  already  adverted  to  the  fact 
that  small-pox  is  highly  contagious,  and  that  contagion  from  those 
suffering  from  it  is  the  means  by  which  the  disease  is  propagated. 

Although  reference  to  infection  appears  in  some  of  the  Arabian  writers,, 
the  contagiousness  of  small-pox  attracted  little  attention  in  this  country 
and  in  western  Europe  until  the  eighteenth  century.  Sydenham  (1624-89), 
though  he  refers  to  the  contagiousness  of  small-pox,  did  not  dwell  upon 
the  matter,  and  did  not  regard  it  as  so  important  an  element  in  the  spread 
of  the  disease  as  some  peculiar  constitution  of  the  atmosphere  to  which 
he  attributed  epidemics.  Boerhaave  was  the  first,  at  the  commencement 
of  the  eighteenth  century,  distinctly  to  formulate  the  now  generally 
accepted  doctrine  that  small-pox  arises  only  from  contagion. 

In  1720,  Mead  drew  up  an  elaborate  system  of  notification,  isolationr 
disinfection,  etc.,  in  view  of  a  threatened  invasion  of  the  plague  ;  but  no 


REPORT   OF  THE   ROYAL   VACCINATION   COMMISSION.  689 

attempt  to  deal  with  small-pox  in  a  similar  fashion  appears  to  have  been, 
made  until  the  last  quarter  of  the  eighteenth  century.  This  was  in  all 
probability  largely  due  to  the  adoption  of  inoculation  as  the  recognised 
defence  against  small-pox,  and  the  acceptance  of  Sydenham's  doctrine 
of  epidemic  causation  may  have  exercised  an  influence  in  the  same 
direction. 

No  writer  appears  to  have  suggested  methods  of  isolation,  disinfection, 
etc.,  against  small-pox  prior  to  1763.  In  that  year  Rast  of  Lyons  published 
his  "  Reflections  on  Inoculation  and  Small-pox,  and  upon  the  means  which . 
might  be  taken  to  deliver  Europe  from  that  malady."  He  maintained— 
(1)  That  small-pox  was  not  a  necessary  and  inevitable  malady  ;  (2)  That  it 
arose  only  from  contagion  ;  (3)  That  it  resembled  plague  in  most  of  its 
features.  His  conclusion  was  expressed  in  these  terms  :  u  I  say,  that  to 
"  deliver  Europe  from  small-pox  we  must  act  upon  principles  directly 
"  opposed  to  inoculation  ;  far  from  multiplying  the  contagion,  we  must 
"  keep  it  away  by  taking  the  same  precautions  and  employing  the  same 
"  measures  against  that  malady  as  have  proved  so  successful  against  leprosy 
"  and  the  plague." 

The  earliest  account  of  the  practical  employment  of  such  means  is 
from  Rhode  Island,  U.S.A.  Haygarth,  on  the  authority  of  Drs.  Moffat  and 
Waterhouse,  states  that  for  many  years  prior  to  1778  small-pox  had  been 
successfully  prevented  from  becoming  epidemic  there  by  regulations  for 
isolation  of  the  infected  on  a  neighbouring  small  island  specially  used  for 
that  purpose,  and  for  quarantining  infected  vessels,  destruction  of  infected 
clothing,  etc.  Moreover,  inoculation  was  discouraged  at  Rhode  Island, 
and  those  who  wished  to  be  inoculated  had  to  go  to  some  place  away  from 
the  Island,  and  were  not  to  return  until  there  was  no  danger  of  their 
infecting  others. 

A  passage  in  Dimsdale's  work  on  Inoculation,  published  in  1781,  shows 
that  in  some  towns  of  England  pest-houses  were  beginning  to  be  used  for 
small-pox.  In  1784  Haygarth,  of  Chester,  published  his  "  Inquiry  how  to 
prevent  the  Small-pox,"  and  in  1793  "A  Sketch  of  a  Plan  to  exterminate 
the  Small-pox  from  Great  Britain." 

The  great  epidemic  of  small-pox  at  Chester  in  1774,  to  which  allusion 
has  already  been  made,  was  the  occasion  of  Haygarth's  first  attempts  at 
organised  dealing  with  epidemics  of  small-pox  with  a  view  to  preven- 
tion. In  his  "  Inquiry  "  he  combated  Sydenham's  doctrine  that  epidemics 
are  due  to  some  occult  condition  of  the  atmosphere,  and  argued  that 
small-pox  was  always  spread  by  infection  only.  He  further  maintained 
that  the  variolous  poison  could  be  carried  as  an  infection  for  a  little 
distance  only  through  the  air,  and  "  consequently  that  the  small-pox  may  be 
"  prevented  by  keeping  persons  liable  to  the  distemper  from  approaching 
"  within  the  infectious  distance  of  the  variolous  poison  till  it  can  be 
"destroyed."  These  views  led  him,  upon  the  return  of  an  epidemic  in 
1777,  to  propose  a  plan  for  the  prevention  of  the  natural  small-pox,  and 
in  1778  a  society  was  formed  to  carry  out  the  plan  in  Chester.  The  plan 
consisted  on  the  one  hand  of  a  general  inoculation  at  people's  homes  at 
stated  intervals,  on  the  ground  that  the  inoculated  small-pox  was  far  less 
fatal  or  injurious  than  the  natural  small-pox,  and  on  the  other  hand  of 

44 


(J90  SUPPLEMENTARY   APPENDIX. 

"  Rules  of  Prevention  "  based  on  Haygarth's  views  of  infection.  In  the 
report  of  the  Society,  called  shortly  uThe  Small-pox  Society,"  dated 
September  1782,  it  is  stated  that  in  the  four  and  a  half  years  of  its 
existence  two  general  inoculations  had  been  held,  and  that  the  deaths 
from  small-pox  had  been  greatly  lessened.  Great  difficulties,  however, 
were  met  with.  "  A  large  proportion  of  the  inhabitants  "  refused  inocula- 
tion, and  a  large  proportion  also,  "  being  fearless,  or  rather  desirous,  that 
their  children  should  be  infected  with  the  natural  small-pox,"  refused  to 
obey  the  Rules  of  Prevention.  Hence,  though  the  same  report  states 
that  the  example  of  Chester  had  been  followed  by  Liverpool,  where 
"  a  general  inoculation  was  successfully  executed  in  the  autumn  of  1781 
and  another  in  the  spring  of  1782,"  and  in  Leeds,  where  a  general 
inoculation  was  held  in  1781  and  another  proposed  in  1782,  with  such 
success  that  the  Royal  College  of  Physicians  in  Edinburgh  appointed  a 
committee  to  inquire  into  "  the  modes  of  conducting  the  general  inocula- 
tions of  the  poor  "  thus  adopted  in  these  places,  the  plan  met  with  such 
difficulties  that  it  was  ultimately  abandoned.  It  will  be  observed  that  a 
general  inoculation  was  an  essential  part  of  the  plan  proposed  and 
carried  out  in  1778-82  ;  but,  writing  in  1784,  Haygarth  looked  forward 
to  being  able  ultimately  to  dispense  with  inoculation,  and  in  the  preface 
to  his  later  edition,  published  in  1793,  he  states  more  definitely  that  the 
adoption  of  his  Rules  of  Prevention  without  any  general  inoculation 
might  exterminate  small-pox  in  some  country  other  than  Great  Britain. 
It  must  be  remembered,  however,  that  Haygarth  entertained  the  opinion 
that  the  infection  of  small-pox  could  not  be  carried  through  the  air  above 
about  half  a  yard,  and  that  no  one  could -be  infected  by  the  clothes  of  a 
person  visiting  a  small-pox  patient  provided  that  he  kept  beyond  this 
distance  from  the  patient.  It  is  obvious  that  if  this  had  been  established 
the  control  of  the  disease  by  isolation  would  be  a  much  simpler  matter 
than  it  really  is. 

In  the  Medico-Chirurgical  Review  for  1796  there  appeared  an  account 
of  a  work  by  Dr.  Faust,  of  Leipsic,  entitled  "  An  Essay  on  the  Duty  of 
a  Man  to  separate  persons  infected  with  the  Small-pox  from  those  in 
"  Health,  thereby  to  effect  the  extirpation  of  that  disease  equally  from 
•"  the  towns  and  countries  of  Europe,"  in  which  it  was  argued  that  the 
first  person  ill  in  a  place  is  the  only  source  from  which  all  the  rest, 
perhaps  hundreds  and  thousands,  become  affected,  and  that  if  he  were 
put  immediately  into  a  situation  where  he  could  not  injure  by  contact 
those  who  had  not  had  the  disorder,  the  spread  of  the  disease  would  be 
prevented. 

In  the  same  Review  for  1799  appeared  an  account  of  establishments 
for  the  extirpation  of  small-pox.  The  failure  of  inoculation  to  attain  the 
desired  end  is  referred  to,  and  legislation  is  urged  to  facilitate  isolation. 
It  is  further  stated  that  in  179(3  the  Prussian  College  of  Physicians  re- 
ported favourably  to  the  King  on  the  project,  and  that  at  Halberstadt  it 
had  been  resolved  to  establish  a  house  for  the  purpose.  At  Cote  d'Or  in 
France  a  similar  plan  had  been  tried  with  success. 

In  1 798  Jenner's  "  Inquiry  "  was  published,  and  in  the  early  years  of  this 
•century  inoculation  began  to  be  discouraged  ;  for  a  while  the  prospects  of 


REPORT   OF   THE   ROYAL   VACCINATION   COMMISSION.  691 

annihilating  small-pox  by  vaccination  appear  to  have  superseded,  in  the 
minds  of  many,  the  plans  of  Haygarth  and  others.  Some  vaccinators, 
however,  like  Willan  and  Ring,  still  looked  to  methods  of  quarantine  and 
to  national  and  municipal  regulations  promoting  isolation  to  exterminate 
the  small-pox. 

It  is  worthy  of  notice,  too,  that  Haygarth  himself,  in  a  letter  quoted 
by  Dr.  Cappe  of  York  in  a  communication  to  the  London  Medical  ami 
Pltyxi&il  Journal  (vol.  iv.,  p.  429),  dated  October  13th,  1800,  remarked, 
"  An  introduction  of  the  vaccine  still  more  than  of  the  variolous  inocula- 
tion would  effectually  promote  the  great  object  of  my  publications/' 

Prior  to  the  year  1866  there  was  no  provision  made  by  law  for  enabling 
sanitary  authorities  to  establish  hospitals  for  infectious  diseases,  and  thus 
to  promote  the  isolation  of  such  cases.  The  only  institutions  of  that 
description  then  existing  were  the  result  of  private  effort.  So  far  as 
regards  small-pox  there  was,  practically  speaking,  no  provision  for  its 
treatment  by  means  of  isolation. 

The  Sanitary  Act  of  1866  empowered,  though  it  did  not  compel, 
local  authorities  throughout  England  and  Wales,  Scotland  and  Ireland, 
to  provide  or  to  join  in  providing  isolation  hospitals  for  the  use  of  the 
inhabitants  of  their  districts.  There  was  further  legislation  on  the 
subject  by  the  Public  Health  Act,  1875  ;  the  Public  Health  (London)  Act, 
1891  ;  the  Public  Health  (Scotland)  Act,  1867  ;  and  the  Public  Health 
(Ireland)  Act,  1878,  into  the  details  of  which  it  is  not  necessary  to  enter. 
The  most  recent  Act  relating  to  the  matter  is  the  Isolation  Hospitals  Act 
of  1893,  which  applies  to  the  small  towns  and  rural  districts  of  England 
and  Wales. 


St(tmj)ing-out  System  in 

Leicester  suffered  severely  from  small-pox  in  1872,  346  deaths  having 
been  registered  as  caused  by  it.  Two  deaths  from  that  disease  occurred 
in  1873,  but  no  other  until  1877,  when  there  were  six,  and  one  in  the 
following  year.  The  next  year  in  which  small-pox  deaths  were  registered 
was  1881.  There  were  two  in  that  year,  and  five  and  three  in  the  follow- 
ing years.  Xo  other  death  took  place  until  1892  and  1893,  in  which 
years  the  fatal  cases  numbered  21. 

Prior  to  1875  the  vaccination  laws  were  well  observed  in  Leicester. 
In  that  year  the  number  of  children  born  who  were  unaccounted  for  \\a< 
only  some  4  per  cent.  Since  then  there  has  been,  as  we  have  seen,  a 
marked  and  progressive  decline  in  the  number  of  vaccinations,  especially 
since  1883,  until  at  the  present  time  80  per  cent,  of  the  children  born 
remain  unvaccinated. 

The  borough  hospital  for  infectious  diseases  was  erected  in  IK  71  -2 
outside  the  town  ;  though  within  the  last  few  years  houses  have  been 
built  in  proximity  to  it.  It  appears  to  have  been  with  Dr.  Crane,  the 
Medical  Officer  of  Health  in  1875,  that  the  quarantining  the  inmates  of 
an  infected  house,  in  addition  to  isolating  the  patient,  originated.  His 
successor,  Dr.  Johnston,  established  it  in  1877  as  a  regular  system.  He 
was  aided  in  this,  after  1879,  by  the  notification  of  infectious  diseases 
then  rendered  compulsory  by  a  private  Act  which  Leicester,  anticipating 


092  SUPPLEMENTARY  APPENDIX. 

most  other  towns,  obtained  in  that  year.  Dr.  Johnston  reported  that  up 
to  1884  the  spread  of  small-pox  from  imported  cases  had  been  arrested  in 
20  instances  by  the  means  he  adopted. 

His  successor,  Dr.  Tomkins,  though,  like  his  predecessors,  regretting 
the  increasing  disuse  of  vaccination,  bore  testimony  in  his  annual  reports* 
to  the  efficacy  of  the  measures  adopted  in  Leicester,  and  expressed  his 
opinion  that  had  such  a  system  been  in  force  at  Sheffield  in  1887  it  would 
not  have  suffered  in  the  way  it  did. 

In  1892  small-pox  became  prevalent  in  different  parts  of  England, 
especially  in  Lancashire  and  Yorkshire.  Many  of  the  large  provincial 
towns  suffered,  and  Leicester  amongst  them.  There  were,  in  1892-3,  357 
cases  of  small-pox  in  Leicester,  of  whom  21,  or  5*8,  died  ;  193  households 
were  invaded,  containing  1234  persons.  The  first  importation  was  by  a 
tramp,  whose  disease,  passing  unrecognised,  caused  infection  at  a  common 
lodging-house  and  at  the  workhouse.  Eleven  other  importations  of  the 
disease  by  tramps  occurred  in  the  course  of  1892-3. 

Leicester  suffered  less  than  many  of  the  other  large  towns  which  have 
been  invaded  by  small-pox  during  recent  years,  both  in  the  number  of 
cases  and  in  the  number  of  deaths.  In  connection  with  this,  however, 
a  point  to  which  we  have  already  called  attention  must  be  borne  in  mind. 
The  disease  was  remarkably  slight  there  in  its  fatality,  even  as  regards 
those  who,  by  reason  of  their  age,  could  not  be  affected  by  the  change  of 
practice  in  relation  to  vaccination.  Dr.  Priestley,  the  Medical  Officer  of 
Health,  claims,  in  his  report  to  the  Sanitary  Committee  for  1893,  that 
it  was  by  reason  of  the  -energetic  methods  adopted  that  the  disease  had 
been  prevented  running  riot  through  the  town.  His  claim  may  be  well 
founded.  At  all  events,  the  experience  of  Leicester  affords  cogent 
evidence  that  the  vigilant  and  prompt  application  of  isolation,  etc.,  even 
with  the  defects  which  were  brought  to  light  during  the  recent  epidemic, 
is  a  most  powerful  agent  in  limiting  the  spread  of  small-pox.  It  is  true 
that  the  system  and  appliances  which  appeared  adequate  for  some  years 
failed  to  prevent  a  serious  outbreak  of  small-pox  in  1892-3.  We  think 
its  value  was  none  the  less  real. 

Stamping-out  System  in  London. 

In  the  Report  of  the  Royal  Commission  of  1881,  already  alluded  to, 
suggestions  were  made  with  regard  to  notification  and  isolation  which 
have  since  been  largely  carried  into  effect.  As  we  have  said,  it  was  con- 
sidered proved  that  the  existing  small-pox  hospitals  had  caused  a  spread 
of  the  disease  in  their  neighbourhood.  We  cannot  but  think  that  this 
may  in  some  measure  account  for  the  greatly  increased  mortality  from 
small-pox  in  London  during  the  1871-72  epidemic  as  compared  with  the 
rest  of  the  country.  It  is  true  that  the  statistics  relating  to  England  and 
Wales  outside  the  Metropolis  include  those  of  other  large  towns  where  the 
same  evil  was  present ;  but  it  probably  did  not  exist  there  in  so  aggravated 
a  form,  and  the  effect  may  be  neutralised  by  the  statistics  relating  to 
smaller  towns  and  rural  districts  with  which  they  are  combined.  This 
idea  has  been  suggested  to  us,  as  the  result  of  the  inquiry,  how  it  has  come 
about  that  whilst  the  Metropolis,  in  the  decennium  1867-76,  and  again 


REPORT   OF   THE    ROYAL   VACCINATION    COMMISSION.  693 

down  to  1885,  compared  so  unfavourably  with  the  rest  of  the  country,  the 
condition  has  since  that  date  become  so  entirely  changed  ?  We  think  it 
is  impossible  to  attribute  this  change  to  vaccination.  There  is  no  reason 
to  suppose  that  the  position  of  the  Metropolis  in  respect  to  vaccination 
has,  since  the  year  1885,  become  superior  to  the  rest  of  England  and 
Wales  :  rather  the  other  way,  as  the  decrease  in  infantile  vaccination  has 
been  greater  during  the  last  few  years  than  in  the  rest  of  England  and 
Wales.  The  change,  therefore,  must  be  due  to  some  other  cause. 

The  hospitals  which,  in  the  opinion  of  the  Commissioners,  were 
propagating  the  disease  in  their  neighbourhood,  were  in  operation 
down  to  July  1882,  when  their  Report  was  made.  In  1877  and  1878,  and 
again  in  1881,  small-pox  was  epidemic  in  London  to  a  considerable  extent. 

We  have  stated  in  detail  in  paragraph  471  *  the  steps  which  were  taken 
by  the  Metropolitan  Asylums  Board  in  consequence  of  the  recommenda- 
tions of  the  Royal  Commission.  It  will  be  seen  that  the  intra-urban 
hospitals  still  continued  in  use,  and  that  complaints  were  made  in  1884  that 
they  were  spreading  small-pox  in  their  vicinity,  although  the  number  in 
each  of  them  was  not  allowed  to  exceed  50.  In  October  1884  this  number 
was  reduced  to  25.  It  was  not,  however,  until  1885  that  the  system  now 
in  operation  was  inaugurated,  and  all  cases  of  small-pox  were  treated  in 
hospital  ships.  It  is  impossible  not  to  be  struck  with  the  fact  that  it  is 
since  the  year  1885  that  the  Metropolis  has  presented  so  satisfactory  an 
aspect  as  regards  small  -pox  mortality.  The  facts  to  which  we  have  been 
calling  attention  certainly  seem  to  point  to  the  conclusion  that  this  has 
been  due  to  a  system  of  isolation,  well  organised  and  administered,  the 
beneficial  effect  of  which  is  no  longer  neutralised  by  a  spread  of  the 
disease  from  the  hospitals  in  which  the  isolation  is  carried  out. 

Upon  the  whole,  we  think  the  experience  of  London  affords  cogent 
evidence  of  the  value  of  a  sound  system  of  isolation  in  checking  the 
spread  of  small-pox. 

Stamping-out  Si/stem  'in   Anstrnlia. 

The  experience  of  isolation  systems  in  Australia  is  interesting  and 
worthy  of  special  notice,  because  whilst  in  this  country  the  quarantining 
of  persons  who  have  come  in  immediate  contact  with  those  suffering  from 
small-pox  has  only  been  possible  with  the  consent  of  the  persons  whom  it 
was  proposed  to  subject  to  quarantine,  in  Australia  their  removal  to  a 
place  of  isolation  has  been  made  compulsory. 

Australia,  by  virtue  of  its  geographical  position,  and  the  consequent 
separation  by  long  sea  voyage  from  infected  ports,  enjoyed  for  a  long 
time  a  sort  of  natural  isolation.  Thus,  Hirsch,  in  his  "  Historical  and 
Geographical  Pathology,"  vol.  i.,  pp.  133-4  (1881),  remarks  :  — 

"  The  continent  of  Australia  up  to  1 838  had  enjoyed  an  absolute 
"  immunity  from  small-pox  ;  towards  the  end  of  that  year  the  disease 
"  appeared  at  Sydney,  having  been  imported  probably  from  China  ;  it 
"  lasted,  however,  only  a  short  time,  and  remained  absent  from  the 
"  continent  until  18G8.  In  that  year  it  was  introduced  into  Melbourne  by 
"  a  ship,  and  again  it  spread  only  to  a  slight  extent,  and  quickly  died  out. 

*  Final  Report. 


694  SUPPLEMENTARY  APPENDIX. 

"  By  a  rigorous  inspection  of  ships  on  their  arrival,  it  has  been  found 
"  possible  to  prevent  subsequent  importations,  a  notable  instance  of  pre- 
"  vention  having  occurred  in  1872.  Tasmania  has  hitherto  quite  escaped 
"  the  disease  ;  so  also  has  New  Zealand,  where  an  importation  of  it  in 
"  1872  was  prevented  by  strictly  isolating  a  vessel  that  had  arrived  with 
'•  small-pox  on  board." 

In  New  South  Wales,  Dr.  MacLaurin,  who  has  been  President  of  the 
Board  of  Health  since  1889,  informed  us  that  the  Government  act  on  the 
assumption  that  small-pox  is  an  exotic  disease,  and  that  every  case  must 
have  come  from  outside  the  colony,  and  it  is  therefore  dealt  with  under 
a  quarantine  Act  of  William  IV.,  originally  instituted  for  dealing  with 
cholera.  By  an  Act  passed  in  1882,  notification  of  small -pox  was  made 
compulsory  on  medical  men  and  householders  under  heavy  penalties.  At 
Sydney  notification  of  small-pox  is  followed  up  by  the  compulsory 
removal  of  the  patient  and  all  persons  who  have  been  in  the  house  with 
the  patient  to  the  quarantine  station  at  North  Head.  This  station  is  670 
acres  in  extent,  and  situated  on  the  peninsula  at  the  mouth  of  Sydney 
Harbour,  and  is  seven  miles  from  the  Health  Office,  with  which  there  is 
telephonic  and  telegraphic  communication.  The  persons  are  conveyed  to 
the  station  by  a  steamboat  comfortably  fitted  expressly  for  the  purpose, 
and  no  difficulty  has  been  experienced  in  effecting  their  removal.  It  was, 
in  Dr.  MacLaurin's  opinion,  by  carrying  out  this  practice  of  isolation  and 
quarantine  that  "  the  epidemic  of  1881-82  was  suppressed,"  and  small-pox 
"  has  'never  become  epidemic  since  this  plan  has  been  adopted."  The 
persons  who  have  been  in  the  house  with  the  patient  are  detained  21  days 
in  quarantine  from  the  date  of  the  last  possible  contagion.  Should  a  case 
of  small-pox  arise  among  them,  those  who  had  been  in  contact  with  such 
infected  person  would  be  detained  for  a  further  period  of  21  days,  and  so 
on.  To  facilitate  this,  the  exposed  persons  are  distributed  in  separate 
groups  within  the  station.  They  are  allowed  to  receive  letters  or  parcels, 
etc.,  and  a  telegraph  operator  is  employed,  "  whose  special  business  it  is 
"  to  work  the  telegraph  at  their  request."  "  Reasonable  compensation  is 
"  given  by  the  Government  for  loss  ;  "  and  there  are  heavy  penalties  under 
the  original  Act  whereby  the  quarantine  is  secured.  The  station  is,  accord- 
ing to  Dr.  MacLaurin,  "  a  pleasant  place  to  stay  in,  and  everything  is  done 
"  that  can  be  done  to  make  the  people  comfortable  :  they  have  nothing 
"  whatever  to  do,  and  are  free  from  all  care,  and  they  can  spend  the  day 
"  pleasantly  enough  ;  but  they  do  not  like  it."  No  one,  however,  raises 
any  objection  to  the  Sydney  system  :  "the  people  are  all  very  sensible 
about  it."  In  all  Australian  towns  the  same  system  is  carried  out  as 
strictly,  with  the  result  that  there  was  not  a  case  of  small-pox  in  Australia 
on  February  5th,  1890  ;  and  Dr.  MacLaurin  is  of  opinion  that  the  risk  of 
dying  of  small-pox  in  Australia  is  smaller  than  in  any  other  part  of  the 
world.  As  regards  vaccination  : — In  New  South  Wales  it  is  very  little 
practised  ;  there  is  no  compulsory  Act  ;  and  though  medical  opinion  is  in 
favour  of  it,  an  opinion  shared  by  Dr.  MacLaurin,  it  is  not  likely  that  a 
compulsory  Vaccination  Act  could  be  passed  or  would  be  tolerated.  The 
proportion  of  young  persons  in  New  South  Wales  who  are  not  vaccinated 
is  accordingly  very  krge  :  probably  much  more  than  half  of  those  under 


REPORT   OF   THE   ROYAL    VACCINATION    COMMISSION.          695 

ten  years  of  age  are  unvaccinated.  Although  Dr.  MacLaurin  favours 
vaccination  and  respects  it  highly,  he  is  satisfied  that  the  system  of  isola- 
tion as  supervised  by  him  is  perfectly  successful.  As  President  of  the 
Board  of  Health  he  considered  it  his  business  to  produce  extinction  of  the 
disease  ;  he  does  not  consider  vaccination  a  sufficiently  absolute  protection 
for  such  purpose  ;  and  he  is  "  fully  of  opinion  that  the  only  way  in  which 
"  you  can  bring  to  an  end  an  outbreak  of  small-pox,  that  is  to  say,  bring 
"  it  under  control,  and  not  leave  it  to  work  itself  out,  is  by  notification 
"  and  isolation.  Of  course,  in  any  small  community,  if  you  let  the  disease 
"  in  it  will  work  itself  out  in  time,  because  all  the  susceptible  people  will 
"  have  had  it ;  but  the  only  way  in  which  you  can  absolutely  control  an 
"  epidemic  of  small-pox  is  by  a  system  of  notification  and  isolation." 

Small-pox  has  never  been  epidemic  in  Western  Australia.  Only  one 
case  has  occurred  within  the  last  31  years,  and  that  was  an  imported  one  ; 
quarantine  was  carried  out,  and  no  infection  occurred  ;  the  immunity  from 
the  disease  is  mainly  at  least  due  to  isolation.  Before  1879  vaccination 
was  not  generally  practised — a  great  majority  of  those  born  in  the  colony 
were  unvaccinated  ;  in  that  year  a  compulsory  Vaccination  Act  was  passed 
in  consequence  of  Sir  H.  Ord  and  Dr.  Waylen's  representations,  and  in 
consequence  of  reports  of  small-pox  in  other  colonies,  and  not  on  account 
of  the  existence  of  small-pox  in  Western  Australia. 

In  Tasmania  there  was  a  compulsory  vaccination  law,  but  it  was  found 
to  be  inoperative  because  no  one  was  appointed  to  conduct  the  prosecu- 
tions, and  it  has  now  fallen  into  desuetude.  The  same  system  of  isolation 
and  quarantine  is  exercised  as  in  the  other  Australian  colonies.  Small-pox 
was  for  the  first  time  introduced  into  Tasmania  in  1887,  and  although 
preparations  for  isolation  were  inadequate,  the  disease  was  soon  stamped 
out.  Communication  between  Launceston  and  Melbourne  was  temporarily 
suspended,  and  to  this  precaution  the  non-invasion  of  Victoria  was 
attributed.  The  particulars  of  this,  the  first  introduction  of  small-pox 
into  Tasmania  during  the  history  of  that  colony,  are  to  be  found  in  a 
report  to  the  Central  Board  of  Health  by  Mr.  A.  Mault,  dated  Novem- 
ber 17th,  1887.  The  origin  of  the  outbreak  is  not  clear,  but  it  was 
presumed  to  have  been  imported,  probably  by  a  ship  from  China,  into 
Launceston.  The  earliest  case  reported  to  the  Local  Board  of  Health 
was  on  September  23rd,  though  it  appears  that  earlier  cases  had  passed 
unnoticed,  or  had  been  notified  as  measles.  Thirty-three  cases  in'  all 
occurred,  every  one  of  which  was  traced  to  direct  infection  from  the  first 
case.  By  September  27th  a  temporary  hospital  had  been  erected,  and 
thither  patients  and  suspects  to  the  number  of  72  were  removed.  The  last 
case  appeared  on  October  13th.  Other  persons  who  had  been  to  the  infected 
houses  were  isolated  in  their  houses  and  watched.  Only  four  of  the  47 
persons  quarantined  at  the  station  were  attacked.  The  clothing  was 
burnt,  and  very  thorough  disinfection  of  the  infected  houses  was  carried 
out,  and  the  dead  were  interred  in  a  special  cemetery.  The  other  colonies 
were  communicated  with,  and  quarantine,  at  first  unduly  rigid  and  after- 
wards relaxed,  was  practised  against  ships  proceeding  from  Tasmania. 
Although  vaccination  had  been  nominally  compulsory  in  Tasmania,  it  was 
estimated  that  two-fifths  of  the  population  were  unvaccinated. 


696  SUPPLEMENTARY  APPENDIX. 


Suggested  Stamping-out  System  In  the  United  Kingdom. 

We  have  no  difficulty  in  answering  the  question,  what  means  other 
than  vaccination  can  be  used  for  diminishing  the  prevalence  of  small-pox  ? 
We  think  that  a  complete  system  of  notification  of  the  disease,  accom- 
panied by  an  immediate  hospital  isolation  of  the  persons  attacked,  together 
with  a  careful  supervision,  or,  if  possible,  isolation  for  sixteen  days  of 
those  who  had  been  in  immediate  contact  with  them,  could  not  but  be 
of  very  high  value  in  diminishing  the  prevalence  of  small-pox.  It  would 
be  necessary,  however,  to  bear  constantly  in  mind,  as  two  conditions  of 
success  :  first,  that  no  considerable  number  of  small-pox  patients  should 
ever  be  kept  together  in  a  hospital  situate  in  a  populous  neighbourhood  : 
and  secondly,  that  the  ambulance  arrangement  should  be  organised  with 
scrupulous  care.  If  these  conditions  were  not  fulfilled,  the  effect  might 
be  to  neutralise  or  even  do  more  than  counteract  the  benefits  otherwise 
flowing  from  a  scheme  of  isolation. 

When  we  turn  to  the  other  branch  of  the  inquiry,  how  far  such  means 
could  be  relied  on  in  the  place  of  vaccination,  we  find  ourselves  involved 
in  questions  of  a  much  more  complicated  nature.  We  have  little  or  no 
experience  to  fall  back  upon.  The  experiment  has  never  been  tried. 
The  nearest  approach  to  a  trial  of  it  has  probably  been  in  Australia.  But 
even  in  the  parts  of  that  country  to  which  we  have  alluded  the  population 
has  not  been  entirely  unvaccinated,  though  there  has  been  a  large 
unvaccinated  class  amongst  it.  Moreover,  in  applying  the  experience  of 
Australia  to  this  country,  two  things  must  be  borne  in  mind.  In  the  first 
place  small-pox  has  only  appeared  from  time  to  time,  introduced  from 
without  at  one  or  other  of  the  ports  of  the  country,  and  the  several 
colonies  of  which  Australia  is  composed  are  of  great  territorial  extent, 
with  few  large  centres  of  population.  In  this  country  small-pox  is 
always  present  in  some  part  of  it.  There  has  not  been  a  single  year 
without  several  deaths  from  the  disease.  Large  centres  of  population 
are  numerous,  and  the  intercourse  between  them  constant.  In  the  several 
colonies  of  Australia  the  number  of  ports  is  not  great,  the  vessels  which 
enter  them  are  comparatively  speaking  not  numerous,  and  the  ports  from 
which  they  arrive  are  many  days'  voyage  distant  ;  and  there  are  careful 
arrangements  for  quarantining  vessels  to  exclude  disease.  The  shipping 
which  enters  English  ports  is  of  vast  quantity,  and  passengers  are  brought 
m  large  numbers  from  the  continent  of  Europe  not  only  daily,  but  it  may 
almost  be  said  hourly  ;  the  voyage,  too,  is  but  brief.  The  other  matter  to 
be  remembered  is,  that  part  of  the  Australian  system  is  the  compulsory 
removal  to  quarantine  for  21  days  of  those  who  have  been  in  the  house 
with  the  patient,  in  addition  to  the  transfer  of  the  patient  himself  to  a 
lospital.  There  can  be  no  doubt  that  such  a  system,  if  completely  carried 
out,  would  be  of  the  highest  efficacy.  But  it  is  obvious  that  in  this 
ountry  the  practical  difficulties  of  working  such  a  scheme  in  the  large 
towns  would  be  really  insuperable,  to  say  nothing  of  the  difficulty  of 
procuring  legislative  sanction  for  it. 


REPORT    OF   THE    ROYAL    VACCINATION    COMMISSION.  697 


Value  <>f 

We  can  see  nothing,  then,  to  warrant  the  conclusion  that  in  this  country 
vaccination  might  safely  be  abandoned,  and  replaced  by  a  system  of 
isolation.  If  such  a  change  were  made  in  our  method  of  dealing  with 
small-pox,  and  that  which  had  been  substituted  for  vaccination  proved 
ineffectual  to  prevent  the  spread  of  the  disease  (it  is  not  suggested  that  it 
could  diminish  its  severity  in  those  attacked),  it  is  impossible  to  contem- 
plate the  consequences  without  dismay. 

To  avoid  misunderstanding,  it  may  be  well  to  repeat  that  we  are  very 
far  from  underrating  the  value  of  a  system  of  isolation.  We  have  already 
dwelt  upon  its  importance.  But  what  it  can  accomplish  as  an  auxiliary 
to  vaccination  is  one  thing  :  whether  it  can  be  relied  on  in  its  stead  is  quite 
another  thing. 

Even  admitting  fully  the  protective  effect  of  vaccination,  it  does  not, 
in  our  opinion,  diminish  the  importance  of  measures  of  isolation  or  dis- 
pense with  their  necessity.  We  think  that  steps  should  be  taken  to  secure 
a  more  general  provision  for  the  isolation  of  small-pox  patients  than 
exists  at  present.  We  have  already  called  attention  to  the  fact  that 
mischievous  results  are  likely  to  follow  the  use  as  a  small-pox  hospital  of 
a  building  situate  in  a  populous  place.  We  think  that  wherever  it  is 
placed  it  should  have  sufficient  space  around  it  to  enable  the  sanitary 
authority  to  add  rapidly  to  the  accommodation  by  the  erection  of  tem- 
porary buildings. 

Compulsory  Pt'oriximt  <>f  Isolation  HojipitcUs. 

Sanitary  authorities  are  now  sometimes  reluctant  to  provide  isolation 
hospitals.  We  think  that,  on  a  petition  by  a  prescribed  number  of  the 
ratepayers  in  a  sanitary  district,  the  Local  Government  Board,  if  satisfied 
that  the  hospital  accommodation  ought  to  be  provided,  should  have  power 
to  make  an  Order  for  such  provision. 


y  Notification  of  >>'/ 

We  think  that  notification  of  small-pox  should  everywhere  be  compul- 
sory, and,  whenever  the  disease  showed  a  tendency  to  become  epidemic, 
a  notice  should  be  served  by  the  sanitary  authority  upon  all  persons  in 
the  neighbourhood  who  would  be  likely  to  come  within  the  reach  of  con- 
tagion, urging  them  to  submit  to  vaccination  or  re-vaccination,  as  the  case 
might  be,  if  they  had  not  been  recently  successfully  vaccinated  or  re- 
vaccinated  ;  and  attention  should  be  called  to  the  facilities  afforded  for 
their  doing  so.  Attention  should  also  be  called  to  the  importance  of 
avoiding  contact  with  persons  suffering  from  the  disease,  or  coming  into 
proximity  to  them,  and  of  avoiding  contact  with  any  person  or  thing 
which  may  have  become  infected.  It  is  important  to  notice  that,  even 
where  vaccination  has  been  neglected,  there  is  great  readiness  to  submit 
to  it  in  the  presence  of  a  threatened  epidemic  ;  a  large  number  of  vacci- 
nations are  then  obtained  willingly  and  without  opposition.  Whenever 


698  SUPPLEMENTARY  APPENDIX. 

a  sanitary  authority  has  received  notification  of  a  case  of  small-poxr 
we  think  the  fact  should  be  at  once  communicated  to  the  vaccination 
authority  of  the  district  in  which  the  case  of  the  disease  has  occurred. 

Regulations  as  to   Tramps,  Inmates  of  Lodging-houses,  etc. 

Our  attention  has  been  drawn  to  the  circumstance  that  outbreaks  of 
small-pox  have  not  unfrequently  had  their  origin  in  the  introduction 
of  the  disease  to  common  lodging-houses  by  tramps  wandering  from  place 
to  place.  In  view  of  this  we  make  the  following  recommendations  : — 

(i.)  That  common  shelters  which  are  not  now  subject  to  the  law 
relating  to  common  lodging-houses  should  be  made  subject 
to  such  law. 

(ii.)  That  there  should  be  power  to  the  local  authority  to  require 
medical  examination  of  all  persons  entering  common  lodging- 
houses  and  casual  wards  to  see  if  they  are  suffering  from 
small-pox,  and  to  offer  a  reward  for  prompt  information  of 
the  presence  of  the  disease. 

(iii.)  That  the  local  authority  should  have  power  to  order  the  keeper 
of  a  common  lodging-house  in  which  there  has  been  small- 
pox to  refuse  fresh  admissions  for  such  time  as  may  be 
required  by  the  authority. 

(iv.)  That  the  local  authority  should  be  empowered  to  require  the 
temporary  closing  of  any  common  lodging-house  in  which 
small-pox  has  occurred. 

(v.)  That  the  local  authority  should  have  power  to  offer  free 
lodgings  to  any  inmate  of  a  common  lodging-house  or  casual 
ward  who  may  reasonably  be  suspected  of  being  liable  to 
convey  small-pox. 

(vi.)  That  the  sanitary  authority  should  give  notice  to  all  adjoining 
sanitary  authorities  of  the  occurrence  of  small-pox  in 
common  lodging-houses  or  casual  wards. 

(vii.)  That  where  the  disease  occurs,  the  Public  Yaccinator  or  the 
Medical  Officer  of  Health  should  attend  and  vaccinate  the 
inmates  of  such  lodging-houses  or  wards,  except  such  as 
should  be  unwilling  to  submit  themselves  to  the  operation. 

Relaxation  of  the  Vaccination  Law. 

After  careful  consideration  and  much  study  of  the  subject,  we  have 

arrived  at  the  conclusion  that  it  would  conduce  to  increased  vaccination 

f  a  scheme  could  be  devised  which  would  preclude  the  attempt  (so  often 

a  vain  one)  to  compel  those  who  are  honestly  opposed  to  the  practice  to 

submit  their  children  to  vaccination,  and,  at  the  same  time,  leave  the  law 

o  operate,  as  at  present,  to  prevent  children  remaining  unvaccinated 

owing  to  the  neglect  or  indifference  of  the  parent.     When  we  speak  of 

honest  opposition  to  the  practice,  we  intend  to  confine  our  remarks  to 

*  m  which  the  objection  is  to  the  operation  itself,  and  to  exclude 


REPORT   OF   THE   ROYAL    VACCINATION    COMMISSION.  699 

cases  in  which  the  objection  arises  merely  from  an  indisposition  to  incur 
the  trouble  involved.  We  do  not  think  such  a  scheme  impossible. 

It  must  of  course  be  a  necessary  condition  of  a  scheme  of  this 
description  that  it  should  be  such  as  would  prevent  an  objection  to  the 
practice  being  alleged  merely  as  an  excuse  to  save  the  trouble  connected 
with  the  vaccination  of  the  child.  We  may  give  the  following  as 
examples  of  the  methods  which  might  be  adopted.  It  might  be  provided 
that  if  a  parent  attended  before  the  local  authority  and  satisfied  them 
that  he  entertained  such  an  objection,  no  proceedings  should  be  taken 
against  him.  Or,  again,  a  statutory  declaration  to  that  effect  before  any 
one  now  authorised  to  take  such  declaration,  or  some  other  specified 
official  or  officials,  might  be  made  a  bar  to  proceedings.  We  do  not  think 
it  would  be  any  real  gain  to  parents  who  had  no  conviction  that  the 
vaccination  of  their  children  was  calculated  to  do  mischief,  to  take  either 
of  these  steps  rather  than  submit  them  to  the  operation. 

It  is  in  England  that  the  point  we  have  been  recently  discussing  is  of 
most  practical  importance,  but  if  our  suggestion  were  adopted  the  change 
should,  of  course,  be  made  in  all  parts  of  the  United  Kingdom. 

(Signed)        HERSCHELL. 

JAMES  PAGET. 

CHARLES  DALRYMPLE. 

W.  GUYER  HUNTER. 

EDWIN  H.  GALSWORTHY. 

JOHN  S.  DUGDALE. 

M.  FOSTER. 

JONATHAN  HUTCHINSON. 

FREDERICK  MEADOWS  WHITE. 

SAM.  WHITBREAD. 

JOHN  A.  BRIGHT. 

BRET  INTK, 
August  1890.  Secretary. 

The  undersigned  do  not  find  themselves  able  to  go  so  far  in  recom- 
mending relaxation  of  the  law  as  is  implied.  We  think  that  in  all  cases- 
in  which  a  parent  or  guardian  refuses  to  allow  vaccination,  the  person  so- 
refusing  should  be  summoned  before  a  magistrate,  as  at  present,  and  that 
the  only  change  made  should  be  to  permit  the  magistrate  to  accept  a 
sworn  deposition  of  conscientious  objection,  and  to  abstain  from  the 
infliction  of  a  fine. 

We  are  also  of  opinion  that,  in  spite  of  the  difficulties  as  set  forth  in 
paragraph  533,*  a  second  vaccination  at  the  age  of  twelve  ought  to  be  mad<* 
compulsory. 

W.  GUYER   HUNTER. 
JONATHAN  HUTCHINSON. 

*  Of  the  Final  RepDrt. 


700  SUPPLEMENTARY  APPENDIX. 

We  the  undersigned  desire  to  express  our  dissent  from  the  proposal  to 
retain  in  any  form  compulsory  vaccination. 

We  cordially  concur  in  the  recommendation  that  conscientious  ob- 
jection to  vaccination  should  be  respected.  The  objection  that  mere 
negligence  or  unwillingness  on  the  part  of  parents  to  take  trouble  might 
keep  many  children  from  being  vaccinated  would  be  largely,  if  not  wholly, 
removed  by  the  adoption  of  the  Scotch  system  of  offering  vaccination  at 
the  home  of  the  child,  and  by  providing  for  medical  treatment  of  any 
untoward  results  which  may  arise. 

We  therefore  think  that  the  modified  form  of  compulsion  recommended 
by  our  colleagues  is  unnecessary,  and  that  in  practice  it  could  not  be 
carried  out. 

The  hostility  which  compulsion  has  evoked  in  the  past  toward  the 
practice  of  vaccination  is  fully  acknowledged  in  the  Report.  In  our 
opinion  the  retention  of  compulsion  in  any  form  will  in  the  future  cause 
irritation  and  hostility  of  the  same  kind. 

The  right  of  the  parent  on  grounds  of  conscience  to  refuse  vaccination 
for  his  child  being  conceded,  and  the  offer  of  vaccination  under  improved 
conditions  being  made  at  the  home  of  the  child,  it  would  in  our  opinion  be 
best  to  leave  the  parent  free  to  accept  or  reject  this  offer. 

SAM.   WHITBREAD. 
JOHN  A.   BRIGHT. 
W.   J.    COLLINS. 
J.  ALLANSON  PICTON. 

Aote. — Dr.  Collins  and  Mr.  Picton  sign  the  above  note  of  reservation, 
tb'itif/h  //„,/  1,,1-re  not  signed  the  Report.  A  statement  of  their  grounds  of 
d'txxi;itf,'oin,  Ike  Report  will  be  found  in  the  form  of  an  Appendix  (65  payei) 
to  the  Report.  They  make  the  following  recommendations  : — 

In  accordance  with  the  sub-head  No.  2  of  the  reference  to  the 
-Commission,  we  would  suggest  the  following  as  the  means  other  than 
vaccination  which  should  be  employed  for  protection  of  a  community 
-from  small-pox  : — 

1.  Prompt  notification  of  any  illness  suspected  to  be  small-pox. 
Improved  instruction  in  the  diagnosis  of  small-pox. 

}.  A  hospital,  suitably  isolated,  of  adequate  accommodation,  in  per- 
manent readiness,  and  capable  of  extension  if  required.  No 
other  disease  to  be  treated  at  the  same  time  in  the  same 
place. 

3.  A  vigilant  sanitary  staff  ready  to  deal  promptly  with  first  cases, 

and  if  necessary  to  make  a  house-to-house  inspection.  The 
medical  officer  of  health  to  receive  such  remuneration  as  to 
render  him  independent  of  private  practice. 

4.  Prompt  removal  to  hospital  by  special   ambulance  of  all  cases 

which  cannot  be  properly  isolated  at  home.  Telephonic  com- 
munication between  Health  Office  and  hospital. 


REPORT   OF  THE   ROYAL   VACCINATION    COMMISSION.  701 

5.  Destruction  of  infected  clothing  and  bedding,  and  thorough  dis- 

infection of  room  or  house  immediately  after  removal  of  the 
patient. 

6.  Daily  observation  (including,  where  possible,  taking  the  tempera- 

ture and  inspection  for  rash)  of  all  persons  who  have  been  in 
close  contact  with  the  patient  during  his  illness  ;  such  super- 
vision to  be  carried  out  either  in  quarantine  stations  (away 
from  the  hospital)  or  at  their  own  homes. 

7.  Closure  of  schools  on  the  occasion  of  the  occurrence  of  small-pox 

among  the  scholars  or  teachers. 

8.  Hospitals  and  quarantine  stations  to  be  comfortable  and  attractive, 

and  so  administered  as  to  secure  the  confidence  of  the  public. 
Hospital  treatment  to  be  free  to  all  classes,  and  compensation 
to  be  paid  to  those  detained  or  otherwise  inconvenienced  in  the 
public  interest,  at  the  public  expense. 

9.  Tramps  entering  casual  wards  to  be  medically  inspected,  their 

clothing  to  be  disinfected,  and  bath  provided.  The  measures 
for  detection  and  isolation  of  small-pox  in  common  lodging- 
houses  suggested  in  section  507  of  the  Report  to  be  carried 
out. 

10.  International  notification  of    the    presence    of    small-pox,   and 

special  vigilance  at  seaports  in  communication  with  infected 
places,  after  the  plan  adopted  in  the  case  of  cholera. 

11.  Attention  to    general   sanitation — prevention   of  overcrowding. 

abundant  water  supply,  and  frequent  removal  of  refuse. 

They  conclude  as  follows  : —    . 

We  believe  the  methods  of  isolation  of  the  infected,  disinfection,  and 
the  observance  of  strict  cleanliness,  are  both  more  successful  and  more 
legitimate  methods  for  the  State  to  encourage.  They  have  the  advantage 
of  applying  the  preventive  only  where  it  is  required  ;  and  they  do  not 
necessitate  an  operation  upon  the  person  of  every  healthy  individual. 

We  therefore  recommend  that  the  law  be  amended  by  the  repeal  of 
the  compulsory  clauses  of  the  Vaccination  Acts.  But  in  consideration 
of  the  prevalent  belief  in  the  value  of  vaccination  as  a  prophylactic  for 
an  indefinite  period,  we  suggest  that  in  other  respects  the  law  should  be 
left  as  it  is,  subject,  however,  to  such  modifications  as  are  recommended 
for  the  diminution  of  attendant  risks.  The  precedent  established  in  the 
case  of  the  abolition  of  compulsory  church  rates  might  be  followed  with 
advantage.  In  that  case  all  machinery  for  laying  and  collecting  the  rate 
was  lef  Antact  though  the  power  of  enforcement  was  taken  away.  The 
effect  of  our  recommendation,  if  adopted,  would  be  that  vaccination 
would  continue  to  be  provided  as  at  present  for  those  who  desire  to  avail 
themselves  of  it,  but  efforts  to  secure  vaccination  would  be  limited  to 
moral  influence— in  a  word,  the  whole  country  would  be  in  the  position  of 
those  unions  in  which  the  guardians  have  abandoned  compulsion. 

The  grounds  on  which  we  object  to  the  enforcement  of  vaccination 
by  penalties  necessarily  lead  us  to  object  to  any  method  of  indirect 


702  SUPPLEMENTARY  APPENDIX. 

compulsion.  We  regard  as  both  inexpedient  and  unjust  exclusion  from 
any  branch  of  the  public  service  because  of  the  refusal  to  submit  to 
vaccination  or  re- vaccination.  The  injustice  is  perhaps  most  severely  felt 
in  the  case  of  candidates  for  employment  as  pupil-teachers  in  public 
elementary  schools.  There  are  now  districts  in  which,  owing  to  the 
general  opposition  to  vaccination,  scarcely  a  girl  or  boy  can  be  found  who 
is  legally  eligible,  and  candidates  have  to  be  brought  in  at  great  incon- 
venience from  surrounding  districts.  The  existence  of  an  exceptional 
case  or  cases  in  which  such  rejected  candidates  have  at  some  time  after- 
wards taken  small-pox  is  in  our  view  no  justification  for  the  continuation 
of  this  grievance.  Statistics  furnished  to  the  Commission  prove  that 
large  numbers  of  vaccinated  or  re-vaccinated  persons  have  taken  the 
disease  ;  and  we  are  not  aware  of  any  evidence  to  show  that  vaccinated 
pupil-teachers  have  any  special  immunity.  If  our  recommendations  were 
carried  out,  the  danger  of  contagion  would  be  greatly  diminished  in 
schools,  as  elsewhere. 

On  the  whole,  then,  while  there  is  much  in  the  report  of  our  colleagues 
from  which  we  dissent,  and  we  have  accordingly  abstained  with  reluctance 
from  adding  our  signatures  to  theirs,  we  are  at  one  with  them  in  holding 
that  it  is  unwise  to  attempt  to  enforce  vaccination  on  those  who  regard  it 
as  useless  and  dangerous.  We,  however,  go  further,  and  agree  with  our 
colleagues,  Mr.  Whitbread  and  Mr.  Bright,  that  it  would  be  simpler  and 
more  logical  to  abolish  compulsory  vaccination  altogether. 


INDEX 


703 


INDEX. 


Abbe's  condenser,  74 
Aberration,  chromatic,  67 

—  spherical,  67 
Abscess,  173 

Achorion  Schonleinii,  584 
Actinomycosis,  413 — 447 

—  bovis,  434 

—  cattle  to  cattle,  444 

—  cultivation  of,  436 
hominis,  431 

• in  cattle,  429 

—  in  man,  426 

transmission    of,   from    man  to 

lower  animals,  443 
Agar-agar,  625 
Air,  compressed,  24 
examination  of,  140 

-  Hesse's  method,  141 

-  Koch's          „       141 

Petri's  „       142 

-  Pouchet's     „       143 

—  Sedgwick's  „       143 
Aitken's  tubes,  129,  630 
Alexines,  57 

Alum  carmine,  616 
Amoeba  coli,  610 
Anaerobes,  classified,  494 

cultivation  of,  130 — 133 

Aniline  water,  617 
Animals  Order,  1878,  455 
Anthrax,  42,  183,  191—216 

-  bacillus  of,  192,  197 

in  horses.  207 

in  swine,  203 

origin  and  spread  of,  198 

preventive  inoculation,  209 

„   measures  in,  199 


Anthrax,  stamping  out  system,  210 
Antiseptics,  30 
Antitoxins,  56,  57 

—  diphtheria,  58 — 62 

septic  infectious,  63 

tetanus,  62 

—  typhoid,  64 
Arthrospores,  19 
Artificial  fluids  as  media,  120 
Ascococcus,  14 

-  Billrothii,  498 

—  citreus,  498 
Ascomycetes,  584 
Asiatic  cholera.  360 
Aspergillus  albus,  588 

clavatus,  588 

flavescens,  588 

—  flavus,  586 

fumigatus,  586 

glaucus,  586 

nidulans,  588 

—  niger,  587 

—  ochraceus,  588 

—  repens,  586 
subfuscus,  588 

B 

Babes'  incubator,  633 
Bacillus,  13,  488 

—  acidiformans,  498 

—  acidi  lactici,  498 

—  aerogenes,  499 
capsulatus,  499 

—  aerophilus.  499 

—  albus,  499 

anaerobiescens,  499 

• cadaveris,  499 

—  putidus,  499 
05  45 


706 


INDEX. 


Bacillus  allantoides,  499 

allii,  499 

alvei,  470 

amylobacter,  502 

amylozyma,  500 

anaerobicus  liquefaciens,  500 

—  anthracis,  192 

aquatilis,  500 

fluorescens,  500 

graveolens,  500 

—  sulcatus,  500 
arborescens,  500 

argenteo-liquefaciens,  501 

—  phosphorescens,  i<0l 

aurantiacus,  501 

aureus,  501 

beroliniensis  indicus,  502 

brassicae,  502 

—  brevis,  502 
brunneus,  502 

buccalis  fortuitus,  502 

—  maximus,  502 
minutus,  502 

butyricus,  502,  503 

cadaveris,  503 

canalis  capsulatus,  504 

—  parvus,  504 

—  candicans,  504 
capsulatus,  504 

mucosus,  504 

—  suis,  504 

—  carabiformis,  504 

—  carnicolor,  504 

—  carotarum,  505 
cavicida,  505 

—  chromo-aromaticus,  505 

Havaniensis,  505 

circulans,  505 

citreus  cadaveris,  505 

cloacag,  505 

coeruleus,  505 

—  coli  communis,  344 

—  similis,  506 

—  constrictus,  506 

—  coprogenes  foetidus,  506 

—  parvus,  506 

—  crassus  aromaticus,  506 

sputigenus,  506 

cuniculicida,  228 

cuticularis,  506 

—  albus,  506 

—  cyaneo-fuscus,  507 


Bacillus  cyaneo-phosphorescens,  507 

cyanogenus,  507,  508 

cystiformis,  508 

—  delicatulus,  508 

dentalis  viridans,  508 

dentriticus,  508 

devorans,  508 

-  diffusus,  508 

-  diphtherias,  332—336 
columbarum,  336 

vitulorum,  509 

dysodes,  509 

endocarditidis  capsulatus,  509- 

griseus,  509 

enteritidis,  372 

epidermidis,  509 

—  erysipelatis  suis,  356 

erythrosporus,  509 

figurans,  510,  511 

filiformis,  511 

Havaniensis,  511 


flavescens,  511 
flavocoriaceus,  511 
fluorescens  aureus,  511 
liquefaciens,  511 


—  longus,  512 

minutissimus,  512 

nivalis,  512 

•  non-liquefaciens,  512 

•  putidus,  512 

—  tenuis,  512 

foetidus,  513 

ozsenae,  513 

—  fulvus,  513 
fuscus,  513 

—  limbatus,  513 
— •  gallinarum,  513 

—  gasoformans,  513 

—  glaucus,  513 

gliscrogenus,  513 

gracilis,  514 

granulosus,  514 

—  graveolens,  514 
guttatus,  514 

haemorrhagic  septicaemia,  231 

—  halophilus,  514 

—  Hansenii,  514 

Havaniensis,  505 

liquefaciens,  514 

—  helvolus,  515 

heminecrobiophilus.  515 

—  hepaticus  fortuitus,  515 


INDEX. 


707 


Bacillus  Hessii,  515 

hyacinth!  septicus,  515 

—  hyalinus,  515 

hydrophilus  fuscus,  515 

ianthinus,  516 

implexus,  516 

—  in  acne  contagiosa  in  horses,  516 
in  cancer,  516 

incanus,  518 

—  in  cholera  in  ducks,  516 

-  in  choleraic  diarrhoea,  516 

—  indicus,  518 
indigogenus,  519 

in  diphtheritic  disease  of  calves, 

516 
--  in  erythema  nodosnm,  517 

in  infectious  disease  of  bees,  471 

inflatus,  519 

in  fowl  enteritis,  230 

in  gangrene,  517 

in  grouse  disease,  517 

in  hog  cholera,  517 

in  infantile  diarrhoea,  517 

in  intestinal  diphtheriain  rabbits, 

517 

in  jequirity  infusion,  517 

in  measles,  517 

in  noma,  517 

in  potato  rot,  517 

in  purpura  heemorrhagica,  517 

in  putrid  bronchitis,  518 

—  in  "red-cod,"  518 

in  rhinoscleroma,  518 

• in  saliva,  518 

—  in  swine  fever,  346-352 
•  erysipelas,  354 

measles,  354 

—  inunctus,  519 
invisibilis,  518 

in  whooping  cough,  518 

—  iridescens,  519 
iris,  519 

lactericeus,  520 

—  lactis  aerogenes,  519 

—  albus,  520 

erythrogenes,  520 

pituitosi,  520 

—  leporis  lethalis,  520 
lepra;,  407 

leptosporus,  520 

—  limbatus  acidi  lactici,  520 
limosus,  520 


Bacillus  liodermos,  521 

liquefaciens,  521 

communis,  521 

magnus,  521 

—  parvus,  521 
liquidus,  521 

-  litoralis,  521 

-  lividus,  524 

-  luteus,  524 

—  maidis,  524 

-  mallei,  452 

megatherium,  524 

—  membranaceus  amethystinus,  525 

meningitidis  purulentae,  525 

mesentericus  f  uscus,  525 

—  ruber,  525 

vulgatus,  526 

inultipediculus,  522 

muscoides,  522 

—  mycoides,  522    fa  ^ 
roseus,  522  ^1» 

—  neapolitanus,  522 
necrophorus,  523 

—  nitrificans,  523 
nodosus  parvus,  523 

—  nubilus,  523 

—  ochraceus,  523 

oedematis  aerobicus,  523 

—  maligni,  220 

—  of  Belfanti  and  Pascarola,  523 

—  of  Colomiatti,  526 

-  of  Fulles,  526 

of  Guillebeau,  526 

of  Letzerich,  526 

—  of  Martinez,  526 

-  of  Nocard,  526 

—  of  Okada,  526 
of  quarter-evil,  217 

—  of  Roth,  526 

-  of  Sattler.  526 

—  of  Schaffer,  527 
of  Scheurleu,  527 

—  of  Schou,  527 

—  of  septicaemia  of  buffaloes,  226 
of  guinea-pigs,  224 

— •  of  mice,  225 

of  swine  plague,  351 

of  Tommasoli,  527 

of  Utpadel,  527 

—  of  Winogradsky,  527 
ophthalmia,  190 

ovatus  minutissimus,  527 


708 


INDEX. 


Bacillus  oxytocus  perniciosus,  527 

pestifer,  528 

phosphorescens  gelidus,  528 

—  indicus,  528 

indigenus,  528 

plicatus,  528 

pneumonias  crouposse,  233 

pneumosepticus,  528 

. polypiformis,  528 

prodigiosus,  528 

proteus  fluorescens,  529 

pseudo-diphtheriticus,  529 

tuberculosis,  529 

—  pulpae  pyogenes,  529 

punctatus,  529 

putrificus  coli,  529 

—  pyocyaneus,  529 

pyogenes  fcetidus,  530 

soli,  530 

—  radiatus,  530 
aquatilis,  530 

ramosus,  531 

reticularis,  531 

rhinoscleroma,  411 

—  rosaceus  metalloides,  531 
rubefaciens,  531 

—  rubellus,  531 

—  ruber,  531 

—  rubescens.  532 
-  rubidus,  532 

sanguinis  typhi,  532 

—  saprogenes,  532 
scissus,  532 

—  septicaemia;  haemorrhagicae,  231 

—  septicus,  532 

acuminatus,  533 

agrigenus,  583 

keratomolaciae,  533 

—  ulceris  gangraenosi,  533 

vesicae,  533 

sessilis,  533 

—  smaragdino-phosphorescens,  533 

—  smaragdtnus  fretidus,  534 
solidus,  534 

—  spiniferus,  534 

spinosus,  534  • 

—  stolonatus,  534 
stolon  if  eras,  534 

—  striatus  albus,  534 

—  flavus,  534 

—  subflavus,  535 
subtilis,  535 


Bacillus  subtilis  similans.  536 

sulfureus,  536 

superficialis,  537 

—  syphilis,  410 

—  tenuis  sputigenus,  537 
termo,  537 

tetani,  457 

thelassopbilus,  537 

thermophilus,  537 

—  tremelloides,  537 

—  tuberculosis,  378 
gallinarum,  402 

—  tumescens,  537 

—  typhi  abdominalis,  342—346 
murium,  359 

—  ubiquitus,  537 

-  ulna,  538 
vacuolosis,  538 

varicosus  conjunctives,  538 

—  venenosus,  538 

brevis,  538 

— . invisibilis,  538 

liquefaciens,  539 

ventriculi,  539 

vermicularis,  539 

vermiculosus,  539 

violaceus,  539 

laurentius,  539 

• virescens,  539 

—  viridis  pallescens,  539 

—  viscosus,  540 

Zurnianus,  540 

Bacteriaceae,  480 

Bacteria,  chemical  action  on,  25 
products  of,  39 

—  composition  of,  11 

—  distribution  of,  29 

-  form  of,  13 
growth  of,  23,  25 

—  in  cattle  plague,  296 

in  diphtheria  of  pigeons,  336 

—  in  distemper,  358 

—  in  foot  and  mouth  disease,  300 
in  horse-pox,  312 

—  in  liquids,  83 

in  louping-ill,  463 

• •  in  measles,  283 

—  in  pus,  176 

—  in  rabies,  460 

in  scarlet  fever,  262 

in  sheep-pox,  298 

—  in  small-pox,  285 


INDEX. 


709 


Bacteria  in  vaccine  lymph,  324—326 

—  in  yellow  fever,  260 

—  microscopic  examination  of,  83 
nitrifying,  27 

—  pathogenic,  27 
saprogenic,  26 

—  stained,  85 

unstained,  84 

Bacterium  aerogenes,  540 

brunneum,  540 

fusiforme,  540 

gingivae  pyogenes,  540 

hyacinth!,  540 

hydrosulfureum  ponticum,  540 

—  litoreum,  540 
luteum,  540 

—  merismopedioides,  541 

navicula,  541 

photometricum,  541 

—  synxanthum,  541 

—  termo,  541 
tholoeideum,  541 

—  ureas,  541 

—  violaceum,  541 

—  Zopfii,  542 
Beggiatoa  alba,  542 

roseo-persicina,  542 

Bibliography,  639 
Bilious  fever,  183 
Biondi's  stain,  615 
Bismarck-brown,  617 
Blepharadenitis,  184 
Blood-serum,  113,  114,  119 
Borax  carmine,  617 
Botrytis  Bassiana,  588 
Bread  paste,  118 
Broncho-pneumonia,  183 
Broth,  118 

Bunge's  method  of  staining  flagella, 
92 


Camera  lucida,  621 
Cancer  bodies,  610 
Caoutchouc  caps,  626 
Caterpillars,  disease  of,  492 
Cattle  plague,  293—296 
—  tuberculosis  of,  :5st) 
Cerebro-spinal  meningitis,  184 
Chemical  action  on  bacteria,  25 

—  disinfectants,  32—36 
products  of  bacteria,  39 


Chemiotaxis,  54 
Chionyphe  Carteri,  588 
Cholera,  360 

—  bacteria  of,  371 

—  comma  bacilli  of,  361 — 367 

—  diarrhoea   from   meat-poisoning, 

371 
— •  diarrhoea  in  fowls,  373 

—  nostras,  370 

—  protective  inoculation,  369 
Ptomaines,  41 

—  spirillum  of,  361 
Chromogenic  bacteria,  25 
Cladothriceas,  480 
Cladothrix,  479 

—  dichotoma,  543 

—  Forsteri,  544 

—  intricata,  544 

—  invulnerabilis,  544 
Classification,  475,  487,  482,  481 
Clostridium  butyricum,  544 

—  fcetidum,  545 
Coccaceaa,  480 
Cocci,  485 
Coccidia,  609 

Comma  bacilli,  361—367 
Cover-glass  preparations,  86 
Cow-pox,  274—282,  312—324,   326  — 

328 
Crenothrix  Kiihniana,  545 

D 

Dacryocystis,  184 
Damp  chambers,  628 
D'Arsonval's  incubator,  631 
Davaine's  septicaemia,  228 
De  Bary's  classification,  481 
Decalcifying,  93,  615 
Defensive  proteids,  57 
Deneke's  comma  bacillus,  367 
Desiccators,  <>::s 
Diarrhoea,  choleraic,  371 
Diphtheria,  330,  1  ^L' 

—  antitoxin  in,  58—62 
-  bacillus  of,  332 

—  in  milk,  338 

in  pigeons,  336 

Loffler's  stain  for,  88 

—  Ptomaines  of,  46 
Diplococcus,  14 

albicans  arnplus,  .">  1 7 

tardissimus.  547 


710 


INDEX. 


Diplococcus     citreus    conglomeratus, 
547 

—  liquefaciens,  547 
coryzae,  547 

flavus  liquefaciens  tardus,  547 

fluorescens  fcetidus,  547 

intercellularis  meningitidis,  548 

•  luteus,  548 

-  pneumonias,  233—238 

—  of  horses,  548 
—  roseus,  548 

subflavus,  548 

Distemper  in  dogs,  358 
Drop-cultures,  120 
Duck  cholera,  230 
Dysentery,  373 

E 

Ebner's  solution,  615 
Egyptian  ophthalmia,  190 
Ehrlich's  staining  method,  89 
Electricity,  24,  127 
Embedding,  93,  615 
Empusa  muscae.  581 

—  radicans,  582 
Endospores,  18 
Enteric,  41,  340 
Enzymes,  48 
Eosin,  89,  617 
Epidemic  disease  of  deer  and  boars, 

227 
of  ferrets,  358 

—  of  mice,  358 
Erysipelas,  185—189 
Esmarch's  roll-cultures,  113 
Experiments  on  animals,  134,  394 


Farrant's  solution,  621 
Favus,  584 

Filter,  hot  water,  624 
Flacherie,  472 
Flagella,  17,  90 
Fliigge's  classification,  481 
Foot  rot,  464 
Form  of  bacteria,  13 
Foul-brood,  469 
Fowl  cholera,  228 
—  enteritis,  230 

scab,  586 

Friedlander's  bacillus,  233 
Fuchsine,  617 


G 

Gangrene,  182 
Gas  chamber,  127 
Gases  produced  by  bacteria,  24 
Gentian  violet  stain,  617 
Giant  cells,  376 
Gibbes'  solution,  618 
Glanders,  451 
— .  bacillus  of,  452 

mallein  in,  454 

• — •—  ptomaine,  48 
Glycerine  agar,  104 
Glycerine  gelatine,  615 
Gonococcus  of  Neisser,  189 
Gonorrhoea,  189 
Gram's  method,  88,  89,  97,  618 
Grease,  303 
Grouse  disease,  230 

H 

Haematococcus  bovis,  548 
Hasmatomonas  carassii,  605 

—  cobitis,  603 
Haematoxylin,  618 
Hgematozoa,  589,  593,  599,  603 
Hardening,  93,  615 
Helicobacterium  aerogenes,  549 
Herpetomonas  Lewisi,  599 
Hessert's  stain,  92 
Historical  introduction,  1 
Horse-pox,  303,  312 

Hot  air  and  steam,  36,  623 
Hot  water  filter,  624 
Hueppe's  classification,  482 

—  inspissator,  630 
Hydrophobia,  459 
Hyphomycetes,  581 
Hypodermii,  581 


Illumination,  75 

Immersion  system,  69 

Immunity,  50 — 57 

Impression  preparations,  92 

Incubators,  631 

Infectious  pleuro-pneumonia,  239,  247 

Influenza,  247 — 249 

Inspissators,  629 

Involution  forms,  15 

Iodine,  618 

Isolation  of  micro-organisms,  139 

Israel's  case,  628 


INDEX. 


711 


K 

Klein's  micrococcus  of  pneumonia,  238 
Kleinenberg's  solution,  616 
Koch's  comma  bacillus,  361 

postulates,  9 

serum  steriliser,  629 

steam  steriliser,  622 


Leprosy,  406 

bacillus  of,  407 

stamping  out  system,  409 

Leptothrix,  480 

• buccalis,  549 

gigantea,  549 

Leuconostoc,  549 
Leukaemia,  184 

Light,  effect  of,  on  bacteria,  24 
Lister's  flasks,  128,  630 
Lithium  carmine  solution,  618 
Lb'ffler's  solution,  619 

—  stain,  88,  90 
Louping  ill,  462 

bacillus  of,  463 

Lutesch's  stain,  91 

M 

Madura  disease,  447 
Magenta  solution,  618 
Malaria,  589 
Malignant  oedema,  220 
Mallein,  454 

Measles,  bacteria  in,  183,  283 
Media,  99 
Merismopedia,  14 
Methyl  violet,  619 
Methylene  blue,  618,  619 
Micrococcus  acidi  lactici,  550 

liquefaciens,  550 

aerogenes,  551 

agilis,  551 

citreus,  551 

—  albus  liquefaciens,  551 
amylivorus,  551 

—  aquatilis,  551 

• invisibilis,  551 

auranticaus,  551 

• botyogenus,  552 

—  candicans,  552 

candidus,  552 

carneus,  552 

cerasinus  siccus,  552 


Micrococcus  cereus  albus,  178 

flavus,  178 

cinnabareus,  552 

—  citreus,  552 

concentricus,  553 

cremoides,  553 

crepusculum,  553 

cumulatus  tenuis,  553 

—  endocarditidis  rugatus,  553 

—  fervidosus,  553 

—  Finlayensis,  553 

—  flavus  desideus,  553 

liquefaciens,  554 

tardigradus,  554 

fcetidus,  554 

Freudenreichi,  554 

—  fuscus,  554 

—  gingivae  pyogenes,  554 

—  gonorrhcese,  190 

—  Havaniensis,  555 

in  abscess  in  rabbits,  556 

in  Biskra  button,  555 

in  gangrenous  mastitis  in  sheep, 

555 

—  in  infectious  pleuro-pneumonia, 

555 

—  in  influenza,  555 
in  pemphigus,  555-6 

-  in  pneumonia,  236,  238,  556 

—  in  py&mia  in  rabbits,  556 

—  in  septicaemia  in  rabbits,  556 

—  in  syphilis,  557 

—  in  trachoma,  190 
lactis  viscosus,  557 

—  luteus,  557 

—  melitensis  (Malta  fever).  557 

—  ochroleucus,  557 

—  plumosus,  557 

—  pneumonias  crouposae,  236 

pyogenes  tenuis,  557 

rosaceus,  558 

—  rosettaceus,  558 

—  salivarus  septicus,  558 

—  stellatus,  588 

—  tetragenus.  558 

mobilis  ventriculi,  558 

subflavus,  558 

versatilis,  558 

—  ureae  liquefaciens,  559 

—  versicolor,  559 

—  violaceus,  559 
viticulosus,  559 


712 


INDEX. 


Micrometer,  80 
Microscope,  65,  612 
Microsporon  furfur,  586 
Microtomes,  94,  614 
Miescher's  tubes,  609 
Mildew,  581 
Milk,  120 

scarlatina,  265,  282 

tubercular,  291 

Miquel's  bulbs,  129 
Moist  chambers  and  cells,  121 
Moitessier's  gas  regulator,  634 
Monads,  601 
Monas  Okenii,  560 
-  Vinosa,  560 

Warmingii,  560 

Mouse  favus,  586 
Movements  of  bacteria,  15,  24 
Mucors,  583 
Muller's  fluid,  616 
Myconostoc  gregarium,  560 
Myco-protein,  11 


Nature  of  soil  for  bacteria,  23 

Neelsen's  solution,  89,  98,  619 

Nicols'  stain,  92 

Nitromonas  of  Winogradsky,  560 

Nutrient  agar,  103 

Nutrient  gelatine,  100 

O 

Oidium  albicans,  586 

lactis,  584 

-  Tuckeri,  584 
Oil  immersion,  70 
Orseille,  619 


Parietti's  method,  148 
Pasteur's  apparatus,  130 
Pathogenic  bacteria,  27 
Pebrine,  471 
Pediococcus  acidi  lactici,  560 

cerevisiae,  560 

Penicillium  glaucum.  588 
Peronospora  infestans,  582 
Petri's  method  exam,  of  air,  142 
Pfeiffer  bodies,  610 
Phagocytosis,  27,  55 
Photogenic  bacteria,  25 
Photography  of  bacteria,  150,  168 


Photo-micrographic  apparatus,  621 

Phycomycetes,  582 

Phylaxins,  57 

Picric  acid,  619 

Picro-carmine,  619 

Picro-lithium  carmine,  620 

Pilobolus,  583 

Plague,  bacillus  of,  250,  252 

Plate  cultivations,  106 

Platinum  needles,  83,  616 

Pleuro-pneumonia,  239 

Pneumo  bacillus     liquefaciens     bovisr 

242,  560 
Pneumonia,  233—238 

ptomaines,  48 

Potash  solution,  620 
Potato  medium,  115 — 117 
Pouchet's  aeroscope,  143 
Preservation  of  preparations,  93 
Protective  inoculation,  49 
-  in  anthrax,  209 

in  cholera,  369 

—  in  rabies,  460 
— -  in  sheep-pox,  298 

in  small-pox,  285,  293 

in  swine  erysipelas,  336 

Proteus  capsulatus  septicus,  560 

—  hominis  capsulatus,  224 

—  in  gangrene  of  the  lung,  560 
—  microsepticus,  560 

-  mirabilis,  561 
— •  septicus,  561 

sulf  ureus,  561 

vulgaris,  561 

Zenkeri,  562 

Pseudo-diphtheritic  bacillus,  335 

Pseudo-diplococcus  pneumonias,  562. 

Psorosperms,  609 

Ptomaines,  39 

Pus,  bacteria  in,  176 

Putrefactive  bacteria,  26,  28 

Pyaemia,  175 


Quarter-evil.  217 


Q 


B 


Eabies,  459—462 
Kag-pickers'  septicaemia,  224. 
Kefraction,  65 
Kelapsing  fever,  257 
Eeproduction,  18 


INDEX. 


via 


Kespiration  of  bacteria,  22 
Rhabdomonas  rosea,  562 
Rhinoscleroma,  411 
Ringworm,  585 

S 
Saccharomyces  acidi  lactici,  580 

albicans,  579 

anomalus,  579 

apiculatus,  578 

aquifolii,  580 

cerevisige,  577 

conglomeratus,  578 

—  ellipsoideus,  577 

exiguus,  578 

glutinus,  579 

Hansenii,  580 

-  ilicis,  580 

Jorgensenii,  578 

Ludwigii,  580 

Marxianus,  580 

membranaefaciens,  580 

minor,  580 

niger,  580 

pastorianus,  578 

pyriformis,  579 

—  rosaceus,  580 

sphsericus,  579 

Safranine,  620 
Sapraemia,  175 
Saprogenic  bacteria,  26 
Saprolegina,  582 
Sapropbytic  bacteria,  27 
Sarcinae,  487 

Sarcina  alba,  562 

—  aurantiaca,  562 

—  Candida,  563 

—  flava,  563 

-  hyalina,  563 
intestinalis,  563 

—  litoralis,  563 

lutea,  563 

mobilis,  563 

—  pulmonum,  563 
Reitenbachii,  564 

—  rosea,  564 

urinae,  5(54 

ventriculi,  564 

Scarlet  fever,  261 
Schizomycetes,  477 

Scblosing's  membrane  regulator,  632 
Sclavo's  stain,  91 


Septicaemia,  175 

—  of  calves,  227 
of  Davaine,  224 

—  of  guinea-pigs,  224 

—  of  mice,  225 
of  rabbits,  228 

—  of  rag-pickers,  224 
Serum  therapy,  56 
Sheep-pox,  297 
Small-pox,  182,  284-293,  326- 
Smut,  581 

Soil,  examination  of,  144    " 

Sozins,  57 

Sphaerotilus  natans,  564 

Spirilla,  495 

Spirillum  amyliferum,  564 

anserum,  564 

— : —  attenuatum,  564 

—  aureum,  564 

—  choleras  Asiaticae,  361 

—  choleroides,  565 
concentricum,  565 

—  dentium,  565 

—  Finkler  and  Prior,  258 
flavescens,  565 

—  flavum,  565 
Giinther,  566 

leucomelaneum,  565 

linguas,  565 

marinum,  565 

— •  Metchnikovi,  373 

— .  MiUer,  566 

—  nasale,  566 
Neisser,  566 

— •  Obermeieri,  258 

—  plicatile,  466 

—  Renon,  566 

—  rosaceum,  566 

—  Rosenbergii,  566 

—  rubrum,  569 

rufum,  566 

rugula,  567 

sanguineum,  567 

—  saprophiles,  567 

—  serpens,  568 

—  Smith,  566 

—  sputigenum,  568 

—  suis,  568 

—  tenue,  568 

tyrogenum,  568 

—  undula,  568 

volutans,  568 


714 


INDEX. 


Spirillum  Weibel,  566 
Spirochcetae,  596 
Spiromonas  Cohnii,  569 

—  volubilis,  569 
Spontaneous  generation,  3 
.Spores,  18 

Staining  spores,  90 
Staphylococcus,  14 

—  cereus  albus,  178,  324 

—  flavus,  178 

—  pyogenes  albus,  178 

—  aureus,  176,  324 

—  citreus,  178 

—  pyosepticus,  569 

—  salivarius  pyogenes,  569 
viridis  flavescens,  569 

Sternberg's  bulbs,  128,  630 
Streptococcus,  14 
acidi  lactici,  569 

—  albus,  569 

—  bombycis,  472 

brevis,  569 

cadaveris,  570 

coli  gracilis,  570 

conglomerates,  570 

—  flavus  desidens,  570 
giganteus  urethrae,  570 

— —  Havaniensis,  570 

—  in  contagious  mammitis  in  cows, 

570 

—  in  progressive  necrosis  in  mice, 

571 

—  in  strangles,  571 

—  liquefaciens,  571 

—  mirabilis,  571 

—  of  Bonome,  571 

—  of  erysipelas,  185,  189 
of  Manneberg,  571 

—  perniciosus  psittacorum,  572 

—  pyogenes,  178-184 

radiatus,  572 

septicus,  572 

—  liquefaciens,  572 

—  vermiformis,  572 
Streptothrix,  496 

—  actinomycotica,  431 

—  albus,  572 

—  asteroides,  573 

—  aurantiaca,  573 
carnea,  573 

—  chromogena,  573 
•  farcinica,  573 


Streptothrix  Forsteri,  573 

—  Hoffmanni,  573 
liquefaciens,  573 

—  Madurae,  449 

musculorum  suis,  573 

odorifera,  573 

violacea,  573 

Substage  condenser,  74 
Suppuration,  48 
Surgical  fever,  182 
Surra,  593 

Swine  erysipelas,  354 
-  fever,  46,  227 

•  measles,  354 

Syphilis,  183,  410 


Tarichium  megaspermum,  582 
Temperature  for  bacteria,  24 
Test-tube  cultivations,  105 
Tetanus,  457 

ptomaine,  41 

Thermo-regulators,  635 
Tilletia  caries,  581 
Toxalbumins,  39 
Trachoma,  412 
Trenkmann's  stain,  91 
Trichomonas,  607 
Tricophyton  tonsurans,  585 
Tripod  levelling  apparatus,  627 
Tuberculosis,  43,  375 

and  meat,  397 

and  public  health,  391 


antitoxin,  64 

-  bacillus  of,  378 
giant  cells  in,  376 

-  in  animals,  389,  394,  399,  400 
in  man,  387 

Typhoid,  340 
Typhus,  259 

U 

Ulcerative  endocarditis,  183 
Undulina  ranarum,  607 
Uredo,  581 

Urine  as  a  medium,  120 
Urobacillus  Duclauxi,  573 
Freudenreichi,  574 

-  Maddoxi,  574 

-  Pasteuri,  574 

—  Schutzenbergi,  574 


INDEX. 


715 


Urocystis  occulta,  581 
Ustilago  carbo,  581 


Van  Ermengen's  stain,  91 
Vegetable  infusions  as  media,  120 
Vesuvin,  620 
Vibrio  rugula,  574 

W 

Warmstage,  123,  124 

Water  bath,  624 

examination  of,  145 


Weigert's  stain,  296 
Wort  gelatine,  104 


;   Xylol,  616 


Yeasts,  577 

Yellow  fever,  182,  259 


Zooglea,  13 

Zopf's  classification,  480 


1896. 


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titioner who  is  beginning  work  in  the  fields  of  ophthalmology,  a  plain 
description  of  the  optical  defects  and  diseases  of  the  eye.  To  this 
end  special  attention  has  been  paid  to  the  clinical  side  of  the  question  ; 
and  the  method  of  examination,  the  symptomatology  leading  to  a 
diagnosis,  and  the  treatment  of  the  various  ocular  defects  have  been 
brought  into  prominence. 

SECOND  EDITION,  REVISED  AND  GREATLY  ENLARGED. 

The  entire  book  has  been  thoroughly  revised.  In  addition  to  this 
general  revision,  special  paragraphs  on  the  following  new  matter  have 
been  introduced  :  Filamentous  Keratitis,  Blood-staining  of  the  Cornea, 
Essential  Phthisis  Bulbi,  Foreign  Bodies  in  the  Lens,  Circinate  Reti- 
nitis,  Symmetrical  Changes  at  the  Macula  Lutea  in  Infancy,  Hyaline 
Bodies  in  the  Papilla,  Monocular  Diplopia,  Subconjunctival  Injections 
of  Germicides,  Infiltration-Anaesthesia,  and  Sterilization  of  Collyria. 
Brief  mention  of  Ophthalmia  Nodosa,  Electric  Ophthalmia,  and  Angioid 
Streaks  in  the  Retina  also  finds  place.  An  Appendix  has  been  added, 
containing  a  full  description  of  the  method  of  determining  the  corneal 
astigmatism  with  the  ophthalmometer  of  Javal  and  Schiotz,  and  the 
rotations  of  the  eyes  with  the  tropometer  of  Stevens.  The  chapter 
on  Operations  has  been  enlarged  and  rewritten. 

A  TEXT-BOOK  OF  HISTOLOGY,  DESCRIPTIVE  AND 
PRACTICAL.  With  174  beautifully  colored  original  figures. 
By  ARTHUR  CLARKSON,  M.  B.,  C.  M.,  Edin.,  formerly  Demon- 
strator of  Physiology  in  the  Owen's  College,  Manchester;  late 
Demonstrator  of  Physiology  in  Yorkshire  College,  Leeds.  Fur- 
nishing the  student  in  one  volume  with  all  that  is  necessary  for 
his  equipment  as  an  Histologist.  Large  8vo.  volume,  strongly 
bound.  Price,  $6.00  net. 

The  purpose  of  the  book  is  to  furnish  the  student  of  Histology,  in 
one  volume,  with  both  the  descriptive  and  the  practical  part  of  the 
science.  In  each  chapter  the  structures  of  the  various  tissues  and 
organs  are  first  described ;  the  student  is  then  taken  tutorially  over  the 
illustrative  specimens;  finally,  an  appendix  concludes  each  chapter 
with  a  short  account  of  the  methods  of  preparation. 


BULLETIN   OF   MEDICAL   PUBLICATIONS:    W.    B.    SAUNDERS,    PHILADA. 


BOOKS -RECENTLY  ISSUED. 


TEXT-BOOK    OF    THE    PATHOGENIC    BACTERIA. 

Specially  written  for  students  of  medicine.  By  JOSEPH  McFAR- 
LAND,  M.  D.,  Professor  of  Pathology  and  Bacteriology  in  the 
Medico- Chirurgical  College  of  Philadelphia,  etc.  359  pages, 
finely  illustrated.  Cloth.  Price,  $2.50  net. 

The  book  presents  a  concise  account  of  the  technical  procedures 
necessary  in  the  study  of  Bacteriology.  It  describes  the  life-history 
of  pathogenic  bacteria,  and  the  pathological  lesions  following  invasions. 
The  illustrations  have  been  gathered  from  standard  sources,  and  com- 
prise the  best  and  most  complete  aggregation  extant. 

"  The  author  has  rendered  a  great  service  to  the  profession  in  bringing  out  this 
work  at  this  opportune  time." — The  American  Therapist,  Aug..  1896. 

PRACTICAL  POINTS  IN  NURSING.  For  Nurses  in 
Private  Practice.  By  EMILY  A.  M.  STONEY,  Superintendent 
of  the  Training-school  for  Nurses,  Carney  Hospital,  South  Boston. 
456  pages,  handsomely  illustrated  with  73  engravings  and  9 
colored  and  half-tone  plates.  i2mo.  Cloth.  Price,  $1.75  net. 

A  vade  mecum  for  the  private  nurse,  and  an  efficient  teaching-book 
for  training-schools.  Instructions  for  quickly  improvising  needed  sick- 
room appliances  constitute  a  valuable  feature  of  the  book.  The 
Appendix  contains,  in  compact  form,  much  information  that  will  be 
of  great  value  to  the  nurse,  including  General  Rules  for  Feeding  the 
Sick ;  Recipes  for  Invalid  Foods  and  Beverages ;  Tables  of  Weights 
and  Measures ;  Table  for  Computing  Date  of  Labor ;  List  of  Abbre- 
viations; Dose-List;  and  a  Glossary  of  64  pages. 

"  The  author's  ^tyle  is  most  agreeable,  and  she  handles  her  subject  in  a  way  that 
clearly  shows  her  complete  familiarity  with  it." — New  York  Polyclinic,  Aug.  15,  1896. 

"  There  are  few  books  intended  for  non-professional  readers  which  can  be  so  cor- 
dially endorsed  by  a  medical  journal  as  can  this  one." — Therapeutic  Gazette,  Aug.  15, 
1896. 

"  This  is  a  well-written,  eminently  practical  volume." — American  Journal  of 
Obstetrics,  Aug.,  1896. 

THE  PICTORIAL  ATLAS  OF  SKIN  DISEASES  AND 
SYPHILITIC  AFFECTIONS  (American  Edition). 
Translation  from  the  French.  Edited  by  J.  J.  PRINGLE,  M.  B., 
F.  R.  C.  P.,  Assistant  Physician  to,  and  Physician  to  the  depart- 
ment for  Diseases  of  the  Skin  at,  the  Middlesex  Hospital,  London. 
Photo-lithochromes  from  the  famous  models  of  dermatological  and 
syphilitic  cases  in  the  Museum  of  the  Saint-Louis  Hospital,  Paris, 
with  explanatory  wood-cuts  and  text.  In  12  Parts,  at  $3.00  per 
Parts  Parts  I.  to  IV.  now  ready. 

"  The  plates  are  beautifully  executed." — JONATHAN  HUTCHINSON,  M.  D.  (London 
Hospital). 


BULLETIN   OF   MEDICAL   PUBLICATIONS:    W.  B.  SAUNDERS,  PHILADA. 


JUST  PUBLISHED. 


A  TEXT-BOOK  OF  BACTERIOLOGY,  including  the  Eti- 
ology and  Prevention  of  Infective  Diseases  and  an  ac- 
count of  Yeasts  and  Moulds,  Hsematozoa,  and  Psoro- 
sperms.  By  EDGAR  M.  CROOKSHANK,  M.  B.,  Professor  of 
Comparative  Pathology  and  Bacteriology,  King's  College,  London. 
A  handsome  octavo  volume  of  700  pages,  illustrated  with  273 
engravings  in  the  text,  and  22  original  and  colored  plates. 
Price,  $6.50  net. 

This  book,  though  nominally  a  Fourth  Edition  of  Professor  Crook- 
shank's  "  MANUAL  OF  BACTERIOLOGY,"  is  practically  a  new  work,  the 
old  one  having  been  reconstructed,  greatly  enlarged,  revised  through- 
out, and  largely  rewritten,  forming  a  text-book  for  the  Bacteriological 
Laboratory,  for  Medical  Officers  of  Health,  and  for  Veterinary  In- 
spectors. 

A  MANUAL  OF  PHYSIOLOGY,  with  Practical  Exer- 
cises. For  Students  and  Practitioners.  By  G.  N.  STEW- 
ART, M.  A.,  M.  D.,  D.  Sc.,  lately  Examiner  in  Physiology, 
University  of  Aberdeen,  and  of  the  New  Museums,  Cambridge 
University;  Professor  of  Physiology  in  the  Western  Reserve 
University,  Cleveland,  Ohio.  Handsome  octavo  volume  of  800 
pages,  with  278  illustrations  in  the  text,  and  5  colored  plates. 
Price,  Cloth,  $3.50  net. 

In  this  book  formal  exposition  has  been  interwoven  with  practical 
work,  in  the  manner  shown  by  the  experience  of  the  Author  to  be 
best  suited  to  the  needs  and  opportunities  of  the  student. 

The  systematic  portion  of  the  book  is  so  arranged  that  it  can  be 
used  independently  of  the  practical  work,  and  aims  at  being  itself  a 
complete  exposition  of  the  subject,  adapted  to  the  requirements  of 
the  student  of  medicine. 

"  It  will  make  its  way  by  sheer  force  of  merit,  and  amply  deserves  tc  do  so.  It  is 
one  of  the  very  best  English  text-books  on  the  subject." — Lancet. 

"Of  the  many  text-books  of  physiology  published,  we  do  not  know  of  one  that  so 
nearly  comes  up  to  the  ideal  as  does  Prof.  Stewart's  volume."—  British  Medical 
Journal. 

WATER  AND  WATER  SUPPLIES.  BY  JOHN  C.  THRESH, 
D.  Sc.,  M.  B.,  D.  P.  H.,  Lecturer  on  Public  Health,  King's  Col- 
lege, London;  Editor  of  the  "Journal  of  State  Medicine,"  etc. 
I2mo,  438  pages,  illustrated.  Handsomely  bound  in  Cloth,  with 
gold  side  and  back  stamps.  Price,  $2.25  net. 

This  work  will  furnish  any  one  interested  in  public  health  the  infor- 
mation requisite  for  forming  an  opinion  as  to  whether  any  supply  or 
proposed  supply  is  sufficiently  wholesome  and  abundant,  and  whether 
the  cost  can  be  considered  reasonable. 


BULLETIN   OF   MEDICAL    PUBLICATIONS  :    W.  B.  SAUNDEUS,  PHILADA. 


JUST  PUBLISHED. 


A  TEXT-BOOK  OF  MATERIA  MEDICA,  THERA- 
PEUTICS, AND  PHARMACOLOGY.  By  GEORGE  F. 
BUTLER,  PH.  G.,  M.  D.,  Professor  of  Materia  Medica  and  of 
Clinical  Medicine  in  the  College  of  Physicians  and  Surgeons, 
Chicago ;  Professor  of  Materia  Medica  and  Therapeutics,  North- 
western University,  Woman's  Medical  School,  etc.  8vo,  858 
pages.  Illustrated.  Prices:  Cloth,  $4.00  net;  Sheep  or  Half- 
Morocco,  $5.00  net. 

A  clear,  concise,  and  practical  text-book,  adapted  for  permanent 
reference  no  less  than  for  the  requirements  of  the  class-room.  The 
arrangement  is  believed  to  be  at  once  the  most  philosophical  and 
rational,  as  well  as  that  best  calculated  to  engage  the  interest  of  those 
to  whom  the  academic  study  of  the  subject  is  wont  to  offer  no  little 
perplexity. 

Special  attention  has  been  given  to  the  Pharmaceutical  section, 
which  is  exceptionally  lucid  and  complete. 

ESSENTIALS  OF  PHYSICAL  DIAGNOSIS  OF  THE 
CHEST.  By  ARTHUR  M.  CORWIN,  A.  M.,  M.  D.,  Demonstrator 
of  Physical  Diagnosis  in  the  Rush  Medical  College,  Chicago; 
Attending  Physician  to  the  Central  Free  Dispensary,  Department 
of  Rhinology,  Laryngology,  and  Diseases  of  the  Chest.  200  pages. 
Illustrated.  Cloth,  flexible  covers.  Price,  $1.25  net. 

ARCHIVES  OF  CLINICAL  SKIAGRAPHY.  By  SYDNEY 
ROWLAND,  B.  A.,  Camb.,  Late  Scholar  of  Downing  College, 
Cambridge,  and  Shuter  Scholar  of  St.  Bartholomew's  Hospital, 
London;  Special  Commissioner  to  "British  Medical  Journal" 
for  the  Investigation  of  the  Applications  of  the  New  Photography 
to  Medicine  and  Surgery.  A  series  of  collotype  illustrations, 
with  descriptive  text,  illustrating  the  applications  of  the  New 
Photography  to  Medicine  and  Surgery.  Price,  per  Part,  $1.00. 
Parts  I.  and  II.  now  ready. 


FORTHCOMING  PUBLICATIONS. 

SURGICAL   DIAGNOSIS   AND   TREATMENT.    By  J. 

W.  MACDONALD,  M.  D.,  Graduate  of  Medicine  of  the  University 
of  Edinburgh  ;  Licentiate  of  the  Royal  College  of  Surgeons  of 
Edinburgh ;  Professor  of  the  Practice  of  Surgery  and  of  Clinical 
Surgery  in  Minneapolis  College  of  Physicians  and  Surgeons; 
Surgeon  to  Wisconsin  Central  Railway,  etc. 

TEXT-BOOK  OF  EMBRYOLOGY.  By  JOHN  C.  HEISLER, 
M.  D.,  Prosector  to  the  Professor  of  Anatomy,  Medical  Depart- 
ment of  the  University  of  Pennsylvania.  , 


BULLETIN    OF    MEDICAL    PUBLICATIONS  :     W.    B.    SAUNDERS,    PHILADA. 


Now  Ready,  Volume  for  1896. 
SAUNDERS' 

American  Year-Book  of  Medicine  and  Surgery 

COLLECTED  AND  ARRANGED  BY  EMINENT  AMERICAN  SPECIALISTS  AND  TEACHERS, 

UNDER    THE    EDITORIAL    CHARGE    OF 

GEORGE  M.  GOULD,  M.  D. 


NOTWITHSTANDING  the  rapid  multiplication  of  medical  and  surgical  works, 
still  these  publications  fail  to  meet  fully  the  requirements  of  the  general  physician, 
inasmuch  as  -he  feels  the  need  of  something  more  than  mere  text-books  of  well- 
known  principles  of  medical  science.  Mr.  Saunders  has  long  been  impressed  with 
this  fact,  which  is  confirmed  by  the  unanimity  of  expression  from  the  profession 
at  large,  as  indicated  by  advices  from  his  large  corps  of  canvassers. 

This  deficiency  would  best  be  met  by  current  journalistic  literature,  but 
most  practitioners  have  scant  access  to  this  almost  unlimited  source  of  informa- 
tion, and  the  busy  practiser  has  but  little  time  to  search  out  in  periodicals  the 
many  interesting  cases,  whose  study  would  doubtless  be  of  inestimable  value  in  his 
practice.  Therefore,  a  work  which  places  before  the  physician  in  convenient  form 
an  epitomization  of  this  literature  by  persons  competent  to  pronounce  upon 

The  Value  of  a  Discovery  or  of  a  Method  of  Treatment 
cannot  but  command  his  highest  appreciation.     It  is   this  critical  and  judicial 
function  that  will  be  assumed  by  the  Editorial  staff  of  the  "  American  Year-Book 
of  Medicine  and  Surgery." 

It  is  the  special  purpose  of  the  Editor,  whose  experience  peculiarly  qualifies 
him  for  the  preparation  of  this  work,  not  only  to  review  the  contributions 
to  American  journals,  but  also  the  methods  and  discoveries  reported  in  the 
leading  medical  journals  of  Europe,  thus  enlarging  the  survey  and  making  the 
work  characteristically  international.  These  reviews  will  not  simply  be  a  series 
of  undigested  abstracts  indiscriminately  run  together,  nor  will  they  be  retro- 
spective of  "news"  one  or  two  years  old,  but  the  treatment  presented  will  be 
synthetic  and  dogmatic,  and  will  include  only  what  is  new.  Moreover,  through 
expert  condensation  by  experienced  writers,  these  discussions  will  be 

COMPRISED  IN  A  SINGLE  VOLUME  OP  ABOUT  1200  PAGES. 

The  work  will  be  replete  with  original  and  selected  illustrations  skilfully 
reproduced,  for  the  most  part,  in  Mr.  Saunders'  own  studios  established  for  the 
purpose,  thus  ensuring  accuracy  in  delineation,  affording  efficient  aids  to  a  right 
comprehension  of  the  text,  and  adding  to  the  attractiveness  of  the  volume. 


Prices:  Cloth,  $6.5O,  net;  Half -Morocco,  $7.5O,  net. 

W.  B.  SAUNDERS,  Publisher, 

925  Walnut  Street,  Philadelphia. 


14  DAY  USE 

RETURN  TO  DESK  FROM  WHICH  BORROWED 

BIOLOGY  LIBRARY 

TEL.  NO.  642-2531 

This  book  is  due  on  the  last  date  stamped  below,  or 

on  the  date  to  which  renewed. 
Renewed  books  are  subject  to  immediate  recall. 


MAR1  -4  1973 


APR-  2  1973 


2 


LD  21A-15m-2,'69 
(J6057slO)476 — A-32 


General  Library 

University  of  California 

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M 

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